Amide-substituted phenyltriazole derivatives and uses thereof

ABSTRACT

The present invention relates to novel 5-(carboxamide)-1-phenyl-1,2,4-triazole derivatives, to processes for the preparation of such compounds, to pharmaceutical compositions containing such compounds, and to the use of such compounds or compositions for the treatment and/or prevention of diseases, in particular for the treatment and/or prevention of renal and cardiovascular diseases.

The present invention relates to novel5-(carboxamide)-1-phenyl-1,2,4-triazole derivatives, to processes forthe preparation of such compounds, to pharmaceutical compositionscontaining such compounds, and to the use of such compounds orcompositions for the treatment and/or prevention of diseases, inparticular for the treatment and/or prevention of renal andcardiovascular diseases.

Vasopressin is a neurohormone which basically regulates waterhomeostasis and vascular tone. It is produced in specialized endocrineneurons in the Nucleus supraopticus and N. paraventricularis in the wallof the third ventricle (hypothalamus) and is transported from therealong the neural processes into the posterior lobes of the hypophysis(neurohypophysis). There, the hormone is released into the bloodstreamin response to different physiological and pathophysiological stimuli. Adisturbed neurohormonal regulation essentially manifests itself in anelevation of the sympathetic tone and inappropriate activation of therenin-angiotensin-aldosterone system (RAAS). While the inhibition ofthese components by beta-receptor blockers on the one hand and by ACEinhibitors or angiotensin-receptor blockers on the other is now aninherent part of the pharmacological treatment of cardiovasculardiseases, the inappropriate elevation of vasopressin secretion is atpresent still not adequately treatable.

Vasopressin exerts its action mainly via binding to three receptors,which are classified as V1a, V1b and V2 receptors and which belong tothe family of G protein-coupled receptors.

V2 receptors are located in the distal tubular epithelium and theepithelium of the collecting tubules in the kidney. Their activationrenders these epithelia permeable to water. This phenomenon is due tothe incorporation of aquaporins (special water channels) in the luminalmembrane of the epithelial cells. Consequently, pharmacologicalinhibition of the action of vasopressin on the V2 receptor results inincreased urine excretion. Hence, drugs with V2 antagonistic activityappear particularly suitable for the treatment of all disease conditionswhich are associated with an overloading of the body with water.

V1b receptors (also named V3 receptors) are mainly detectable in thecentral nervous system. Together with corticotropin-releasing hormone(CRH), vasopressin regulates the basal and stressinduced secretion ofadrenocorticotropic hormone (ACTH) via the V1b receptor.

V1a receptors are mainly located on vascular smooth muscle cells (VSMC)but also on cardiomyocytes, fibroblasts and specialized renal cells likeglomerular mesangial cells or cells of the macula densa which controlthe release of renin [Wasilewski M A, Myers V D, Recchia F A, Feldman AM, Tilley D G, Cell Signal., 28(3), 224-233, (2016)]. The activation ofVSMC V1a receptor by vasopressin gives rise to intracellular calciumrelease and according vasoconstriction. Therefore, stimulation of VSMCV1a receptors causes increased vascular resistance and increased cardiacafterload. Cardiac output is adversely affected by V1a-mediatedvasoconstriction. The increase in afterload and direct stimulation ofV1a receptors on cardiomyocytes can lead to cardiac hypertrophy andremodeling including fibrosis. Mice with cardiac-specific overexpressionof V1a receptor develop cardiac hypertrophy leading to dilation and leftventricular dysfunction, suggesting an essential role for V1a receptorin the development of heart failure [Li X, Chan T O, Myers V, ChowdhuryI, Zhang X Q, Song J, Zhang J, Andrel J, Funakoshi H, Robbins J, Koch WJ, Hyslop T, Cheung J Y, Feldman A M, Circulation.; 124, 572-581(2011)].

V1a receptor is also expressed in the renal cortical and medullaryvasculature, where it mediates vasoconstriction of renal vessels andaffecting overall renal blood flow. Thus, the activation of V1a receptorcan decrease renal medullary blood flow inducing further pathologicalprocesses as tissue hypoxia, reduced oxygen and accordingly energysupply for tubular transport processes as well as direct damages ofmesangial and macula densa cells. It has been demonstrated thatmesangial V1a receptor activation mediates TGFβ signaling and causes anincrease in production of collagen IV. While this signaling contributesto extracellular matrix accumulation and remodeling in the kidney,similar signaling pathways are believed to occur in cardiac cellsespecially after myocardial infarction, which emphasizes the centralrole of V1a receptor in the development of hypertrophic and fibroticprocesses in response to pathophysiological elevated vasopressin levels[Wasilewski M A, Myers V D, Recchia F A, Feldman A M, Tilley D G.Arginine vasopressin receptor signaling and functional outcomes in heartfailure. Cell Signal., 28(3), 224-233 (2016)].

Since V1a receptors are mainly expressed on VSMCs and thus participatingin vascular function, a link to vascular diseases as peripheral arterialdisease (PAD) including claudication and critical limb ischemia as wellas coronary microvascular dysfunction (CMD) is conceivable.

Apart from this, V1a receptors are also expressed on human platelets andin the liver. The meaning of platelet V1a receptors is not fullyunderstood although vasopressin induces aggregation of human plateletsvia V1a receptor at high concentrations ex vivo. Therefore, inhibitionof vasopressin-induced platelet aggregation by V1a receptor antagonistsis a useful pharmacological ex vivo assay making use of human tissueendogenously expressing the V1a receptor [Thibonnier M, Roberts J M, JClin Invest.; 76:1857-1864, (1985)].

Vasopressin stimulates gluconeogenesis and glycogenolysis via activationof the hepatic V1a receptor. Animal studies have shown that vasopressinimpairs glucose tolerance which could be inhibited by a V1a receptorantagonist thereby providing a link of vasopressin receptor V1a todiabetes mellitus. [Taveau C, Chollet C, Waeckel L, Desposito D, BichetD G, Arthus M F, Magnan C, Philippe E, Paradis V, Foufelle F, HainaultI, Enhorning S, Velho G, Roussel R, Bankir L, Melander O, Bouby N.Vasopressin and hydration play a major role in the development ofglucose intolerance and hepatic steatosis in obese rats. Diabetologia.,58(5), 1081-1090, (2015)]. Vasopressin was shown to contribute to thedevelopment of albuminuria and to diabetes-induced nephropathy in animalmodels which is consistent with epidemiological findings in humans.

It was found recently that vasopressin also seems to play a causal rolein the development of preeclampsia. Chronic infusion of vasopressinduring pregnancy in mice is sufficient to induce all of the majormaternal and fetal phenotypes associated with human preeclampsia,including pregnancy-specific hypertension [Santillan M K, Santillan D A,Scroggins S M, Min J Y, Sandgren J A, Pearson N A, Leslie K K, Hunter SK, Zamba G K, Gibson-Corley K N, Grobe J L. Vasopressin in preeclampsia:a novel very early human pregnancy biomarker and clinically relevantmouse model. Hypertension. 64(4), 852-859, (2014)].

Vasopressin levels can be elevated in women with dysmenorrhoea (agynecological disorder which is characterised by cyclical crampingpelvic pain) during menstruation, which appear to increase myometrialsmooth muscle contraction. It was found recently that a selectivevasopressin V1a receptor antagonist (relcovaptan/SR-49059) can reduceintrauterine contractions elicited by vasopressin.

For these reasons, agents which inhibit the action of vasopressin on theV1a receptor appear suitable for the treatment of several cardiovasculardiseases. In particular, agents which inhibit the action of vasopressinselectively on the V1a receptor offer an especially ideal profile forthe treatment of otherwise normovolemic patients, i.e. those which arenot eligible for decongestion by e.g. high doses of loop diuretics or V2antagonists, and where induced aquaresis via V2 inhibition may beundesired.

Certain 4-phenyl-1,2,4-triazol-3-yl derivatives have been described inWO 2005/063754-A1 and WO 2005/105779-A1 to act as vasopressin V1areceptor antagonists that are useful for the treatment of gynecologicaldisorders, notably menstrual disorders such as dysmenorrhea.

In WO 2011/104322-A1, a particular group of bis-aryl-bonded1,2,4-triazol-3-ones, including 5 phenyl-1,2,4-triazol-3-yl and1-phenyl-1,2,3-triazol-4-yl derivatives thereof, has been disclosed asantagonists of vasopressin V2 and/or V1a receptors being useful for thetreatment and/or prevention of cardiovascular diseases. The describedcompounds, however, do not show sufficient selectivity towards the V1areceptor and mostly show combined activity on both vasopressin V1a andV2 receptors. Yet, as outlined above, a high affinity as well asselectivity for the V1a receptor is a desirable prerequisite for thetreatment of disease conditions where a decongestion is not desired andmay lead to a dysregulated body fluid homeostasis including decreasedblood plasma osmolality in otherwise normovolemic individuals.

In WO 2016/071212-A1 certain 5-(hydroxyalkyl)-1-phenyl-1,2,4-triazolederivatives have been disclosed, which act as potent antagonists of bothvasopressin V1a and V2 receptors and, in addition, exhibit significantlyenhanced aquaretic potency in vivo after oral application. The compoundsare described to be useful for the treatment and/or prevention ofcardiovascular and renal diseases. Yet, as outlined above, a highaffinity as well as selectivity for the V1a receptor is a desirableprerequisite for the treatment of disease conditions where adecongestion is not desired and may lead to a dysregulated body fluidhomeostasis including decreased blood plasma osmolality in otherwisenormovolemic individuals.

An activity profile with a high selectivity for the V1a receptor has alow potential to cause unwanted off-target related side effects andwould also help towards reducing the amount of substance which is goingto be required to achieve and maintain the desired therapeutic effect,thus limiting the potential for unacceptable side effects and/orunwanted drug-drug interactions during the treatment of patients whichmight already be at high risk, such as, for example, in acute or chronicheart and kidney diseases.

The technical problem to be solved according to the present inventionmay therefore be seen in identifying and providing new compounds thatact as potent antagonists of the vasopressin V1a receptor. A furtherobject of the invention is to identify and provide new compounds with ahigh affinity and selectivity vis-h-vis the vasopressin V1a receptor.The compounds are intended to avoid inducing aquaresis via V2inhibition. The compounds are further intended to have a similar orimproved therapeutic profile compared to the compounds known from theprior art, for example with respect to their in vivo properties, forexample their pharmacokinetic and pharmacodynamic characteristics and/ortheir metabolic profile and/or their dose-activity relationship.

Surprisingly, it has now been found that certain5-(carboxamide)-1-phenyl-1,2,4-triazole derivatives represent highlypotent and selective antagonists of the V1a receptor. This specificprofile renders the compounds of the present invention useful for thetreatment and/or prevention of diseases, which are associated with V1areceptor activation. The compounds of the present invention areparticularly useful for the treatment and/or prevention of renal andcardiovascular diseases in subjects which do not suffer from fluidoverload and who therefore should not be decongested.

The compounds of the present invention have valuable pharmacologicalproperties and can be used for the prevention and/or treatment ofvarious diseases and disease-induced states in humans and other mammals.

In one aspect, the present invention relates to5-(carboxamide)-1-phenyl-1,2,4-triazole derivatives of the generalformula (I)

-   -   in which    -   R¹ represents a group of the formula

-   -   -   in which        -   #¹ represents the point of attachment to the nitrogen atom,

    -   Ar represents a group of the formula

-   -   -   in which        -   #² represents the point of attachment to the nitrogen atom,        -   R² represents a group selected from trifluoromethyl,            trifluoromethoxy, ethoxy, —C(═O)N(H)CH₃ and —S(═O)₂CH₃.

The compounds according to this invention can also be present in theform of their salts, solvates and/or solvates of the salts.

The term “comprising” when used in the specification includes“consisting of”.

If within the present text any item is referred to as “as mentionedherein”, it means that it may be mentioned anywhere in the present text.

The terms as mentioned in the present text have the following meanings:

The term “C₁-C₄-alkyl” means a linear or branched, saturated, monovalenthydrocarbon group having 1, 2, 3, or 4 carbon atoms, e.g. a methyl,ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, or anisomer thereof. Particularly, said group has 1, 2, 3 or 4 carbon atoms(“C₁-C₄-alkyl”), e.g. a methyl, ethyl, propyl, isopropyl, butyl,sec-butyl isobutyl, or tert-butyl group, more particularly 1, 2 or 3carbon atoms (“C₁-C₃-alkyl”), e.g. a methyl, ethyl, n-propyl orisopropyl group, even more particularly a methyl group.

It is possible for the compounds of general formula (I) to exist asisotopic variants. The invention therefore includes one or more isotopicvariant(s) of the compounds of general formula (I), particularlydeuterium-containing compounds of general formula (I).

The term “Isotopic variant” of a compound or a reagent is defined as acompound exhibiting an unnatural proportion of one or more of theisotopes that constitute such a compound.

The term “Isotopic variant of the compound of general formula (I)” isdefined as a compound of general formula (I) exhibiting an unnaturalproportion of one or more of the isotopes that constitute such acompound.

The expression “unnatural proportion” means a proportion of such isotopewhich is higher than its natural abundance. The natural abundances ofisotopes to be applied in this context are described in “IsotopicCompositions of the Elements 1997”, Pure Appl. Chem., 70(1), 217-235,1998.

Examples of such isotopes include stable and radioactive isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine,chlorine, bromine and iodine, such as ²H (deuterium), ³H (tritium), ¹¹C,¹³C, ¹⁴C, ¹⁵N, ¹⁷O, ¹⁸O, ³²P, ³³P, ³³S, ³⁴S, ³⁵S, ³⁶S, ¹⁸F, ³⁶Cl, ⁸²Br,¹²³I, ¹²⁴I, ¹²⁵I, ¹²⁹I and ¹³¹I, respectively.

With respect to the treatment and/or prevention of the disordersspecified herein the isotopic variant(s) of the compounds of generalformula (I) preferably contain deuterium (“deuterium-containingcompounds of general formula (I)”). Isotopic variants of the compoundsof general formula (I) in which one or more radioactive isotopes, suchas ³H or ¹⁴C, are incorporated are useful e.g. in drug and/or substratetissue distribution studies. These isotopes are particularly preferredfor the ease of their incorporation and detectability. Positron emittingisotopes such as ¹⁸F or ¹¹C may be incorporated into a compound ofgeneral formula (I). These isotopic variants of the compounds of generalformula (I) are useful for in vivo imaging applications.Deuterium-containing and ¹³C-containing compounds of general formula (I)can be used in mass spectrometry analyses (H. J. Leis et al., Curr. Org.Chem., 1998, 2, 131) in the context of preclinical or clinical studies.

Isotopic variants of the compounds of general formula (I) can generallybe prepared by methods known to a person skilled in the art, such asthose described in the schemes and/or examples herein, by substituting areagent for an isotopic variant of said reagent, preferably for adeuterium-containing reagent. Depending on the desired sites ofdeuteration, in some cases deuterium from D₂O can be incorporated eitherdirectly into the compounds or into reagents that are useful forsynthesizing such compounds (Esaki et al., Tetrahedron, 2006, 62, 10954;Esaki et al., Chem. Eur. J., 2007, 13, 4052). Deuterium gas is also auseful reagent for incorporating deuterium into molecules. Catalyticdeuteration of olefinic bonds (H. J. Leis et al., Curr. Org. Chem.,1998, 2, 131; J. R. Morandi et al., J. Org. Chem., 1969, 34 (6), 1889)and acetylenic bonds (N. H. Khan, J. Am. Chem. Soc., 1952, 74 (12),3018; S. Chandrasekhar et al., Tetrahedron Letters, 2011, 52, 3865) is adirect route for incorporation of deuterium. Metal catalysts (i.e. Pd,Pt, and Rh) in the presence of deuterium gas can be used to directlyexchange deuterium for hydrogen in functional groups containinghydrocarbons (J. G. Atkinson et al., U.S. Pat. No. 3,966,781). A varietyof deuterated reagents and synthetic building blocks are commerciallyavailable from companies such as for example C/D/N Isotopes, Quebec,Canada; Cambridge Isotope Laboratories Inc., Andover, Mass., USA; andCombiPhos Catalysts, Inc., Princeton, N.J., USA. Further information onthe state of the art with respect to deuterium-hydrogen exchange isgiven for example in Hanzlik et al., J. Org. Chem. 55, 3992-3997, 1990;R. P. Hanzlik et al., Biochem. Biophys. Res. Commun. 160, 844, 1989; P.J. Reider et al., J. Org. Chem. 52, 3326-3334, 1987; M. Jarman et al.,Carcinogenesis 16(4), 683-688, 1995; J. Atzrodt et al., Angew. Chem.,Int. Ed. 2007, 46, 7744; K. Matoishi et al., Chem. Commun. 2000,1519-1520; K. Kassahun et al., WO2012/112363.

The term “deuterium-containing compound of general formula (I)” isdefined as a compound of general formula (I), in which one or morehydrogen atom(s) is/are replaced by one or more deuterium atom(s) and inwhich the abundance of deuterium at each deuterated position of thecompound of general formula (I) is higher than the natural abundance ofdeuterium, which is about 0.015%. Particularly, in adeuterium-containing compound of general formula (I) the abundance ofdeuterium at each deuterated position of the compound of general formula(I) is higher than 10%, 20%, 30%, 40%, 50%, 60%, 70% or 80%, preferablyhigher than 90%, 95%, 96% or 97%, even more preferably higher than 98%or 99% at said position(s). It is understood that the abundance ofdeuterium at each deuterated position is independent of the abundance ofdeuterium at other deuterated position(s).

The selective incorporation of one or more deuterium atom(s) into acompound of general formula (I) may alter the physicochemical properties(such as for example acidity [C. L. Perrin, et al., J. Am. Chem. Soc.,2007, 129, 4490; A. Streitwieser et al., J. Am. Chem. Soc., 1963, 85,2759;], basicity [C. L. Perrin et al., J. Am. Chem. Soc., 2005, 127,9641; C. L. Perrin, et al., J. Am. Chem. Soc., 2003, 125, 15008; C. L.Perrin in Advances in Physical Organic Chemistry, 44, 144],lipophilicity [B. Testa et al., Int. J. Pharm., 1984, 19(3), 271])and/or the metabolic profile of the molecule and may result in changesin the ratio of parent compound to metabolites or in the amounts ofmetabolites formed. Such changes may result in certain therapeuticadvantages and hence may be preferred in some circumstances. Reducedrates of metabolism and metabolic switching, where the ratio ofmetabolites is changed, have been reported (A. E. Mutlib et al.,Toxicol. Appl. Pharmacol., 2000, 169, 102; D. J. Kushner et al., Can. J.Physiol. Pharmacol., 1999, 77, 79). These changes in the exposure toparent drug and metabolites can have important consequences with respectto the pharmacodynamics, tolerability and efficacy of adeuterium-containing compound of general formula (I). In some casesdeuterium substitution reduces or eliminates the formation of anundesired or toxic metabolite and enhances the formation of a desiredmetabolite (e.g. Nevirapine: A. M. Sharma et al., Chem. Res. Toxicol.,2013, 26, 410; Efavirenz: A. E. Mutlib et al., Toxicol. Appl.Pharmacol., 2000, 169, 102). In other cases the major effect ofdeuteration is to reduce the rate of systemic clearance. As a result,the biological half-life of the compound is increased. The potentialclinical benefits would include the ability to maintain similar systemicexposure with decreased peak levels and increased trough levels. Thiscould result in lower side effects and enhanced efficacy, depending onthe particular compound's pharmacokinetic/pharmacodynamic relationship.ML-337 (C. J. Wenthur et al., J. Med. Chem., 2013, 56, 5208) andOdanacatib (K. Kassahun et al., WO2012/112363) are examples for thisdeuterium effect. Still other cases have been reported in which reducedrates of metabolism result in an increase in exposure of the drugwithout changing the rate of systemic clearance (e.g. Rofecoxib: F.Schneider et al., Arzneim. Forsch./Drug. Res., 2006, 56, 295;Telaprevir: F. Maltais et al., J. Med. Chem., 2009, 52, 7993).Deuterated drugs showing this effect may have reduced dosingrequirements (e.g. lower number of doses or lower dosage to achieve thedesired effect) and/or may produce lower metabolite loads.

A compound of general formula (I) may have multiple potential sites ofattack for metabolism. To optimize the above-described effects onphysicochemical properties and metabolic profile, deuterium-containingcompounds of general formula (I) having a certain pattern of one or moredeuterium-hydrogen exchange(s) can be selected. Particularly, thedeuterium atom(s) of deuterium-containing compound(s) of general formula(I) is/are attached to a carbon atom and/or is/are located at thosepositions of the compound of general formula (I), which are sites ofattack for metabolizing enzymes such as e.g. cytochrome P₄₅₀.

Where the plural form of the word compounds, salts, polymorphs,hydrates, solvates and the like, is used herein, this is taken to meanalso a single compound, salt, polymorph, isomer, hydrate, solvate or thelike.

By “stable compound” or “stable structure” is meant a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

The compounds of the present invention optionally contain one asymmetriccentre, depending upon the location and nature of the varioussubstituents desired. It is possible that one asymmetric carbon atom ispresent in the (R) or (S) configuration, which can result in racemicmixtures. In certain instances, it is possible that asymmetry also bepresent due to restricted rotation about a given bond, for example, thecentral bond adjoining two substituted aromatic rings of the specifiedcompounds. Preferred compounds are those which produce the moredesirable biological activity. Separated, pure or partially purifiedisomers and stereoisomers or racemic mixtures of the compounds of thepresent invention are also included within the scope of the presentinvention. The purification and the separation of such materials can beaccomplished by standard techniques known in the art.

The optical isomers can be obtained by resolution of the racemicmixtures according to conventional processes, for example, by theformation of diastereoisomeric salts using an optically active acid orbase or formation of covalent diastereomers. Examples of appropriateacids are tartaric, diacetyltartaric, ditoluoyltartaric andcamphorsulfonic acid. Mixtures of diastereoisomers can be separated intotheir individual diastereomers on the basis of their physical and/orchemical differences by methods known in the art, for example, bychromatography or fractional crystallisation. The optically active basesor acids are then liberated from the separated diastereomeric salts. Adifferent process for separation of optical isomers involves the use ofchiral chromatography (e.g., HPLC columns using a chiral phase), with orwithout conventional derivatisation, optimally chosen to maximise theseparation of the enantiomers. Suitable HPLC columns using a chiralphase are commercially available, such as those manufactured by Daicel,e.g., Chiracel O D and Chiracel O J, for example, among many others,which are all routinely selectable. Enzymatic separations, with orwithout derivatisation, are also useful. The optically active compoundsof the present invention can likewise be obtained by chiral synthesesutilizing optically active starting materials. In order to distinguishdifferent types of isomers from each other reference is made to IUPACRules Section E (Pure Appl Chem 45, 11-30, 1976).

The present invention includes all possible stereoisomers of thecompounds of the present invention as single stereoisomers, or as anymixture of said stereoisomers, e.g. (R)- or (S)-isomers, in any ratio.Isolation of a single stereoisomer, e.g. a single enantiomer or a singlediastereomer, of a compound of the present invention is achieved by anysuitable state of the art method, such as chromatography, especiallychiral chromatography, for example.

Further, it is possible for the compounds of the present invention toexist as tautomers. The present invention includes all possibletautomers of the compounds of the present invention as single tautomers,or as any mixture of said tautomers, in any ratio.

Further, the compounds of the present invention can exist as N-oxides,which are defined in that at least one nitrogen of the compounds of thepresent invention is oxidised. The present invention includes all suchpossible N-oxides.

The present invention also covers useful forms of the compounds of thepresent invention, such as metabolites, hydrates, solvates, salts, inparticular pharmaceutically acceptable salts, and/or co-precipitates.

The compounds of the present invention can exist as a hydrate, or as asolvate, wherein the compounds of the present invention contain polarsolvents, in particular water, methanol or ethanol for example, asstructural element of the crystal lattice of the compounds. It ispossible for the amount of polar solvents, in particular water, to existin a stoichiometric or non-stoichiometric ratio. In the case ofstoichiometric solvates, e.g. a hydrate, hemi-, (semi-), mono-, sesqui-,di-, tri-, tetra-, penta-etc. solvates or hydrates, respectively, arepossible. The present invention includes all such hydrates or solvates.Hydrates are preferred solvates in the context of the present invention.

In particular, the 3,3,3-trifluoro-2-oxopropyl derivatives of theformula (I-B) according to the invention (ketone form) may also bepresent in the 3,3,3-trifluoro-2,2-dihydroxypropyl form (I-B)′ (hydrateform) (see Scheme 1 below); both forms are expressly embraced by thepresent invention.

Further, it is possible for the compounds of the present invention toexist in free form, e.g. as a free base, or as a free acid, or as azwitterion, or to exist in the form of a salt. Said salt may be anysalt, either an organic or inorganic addition salt, particularly anypharmaceutically acceptable organic or inorganic addition salt, which iscustomarily used in pharmacy, or which is used, for example, forisolating or purifying the compounds of the present invention.

The term “pharmaceutically acceptable salt” refers to an inorganic ororganic acid addition salt of a compound of the present invention. Forexample, see S. M. Berge, et al. “Pharmaceutical Salts,” J. Pharm. Sci.1977, 66, 1-19.

A suitable pharmaceutically acceptable salt of the compounds of thepresent invention may be, for example, an acid-addition salt of acompound of the present invention bearing a nitrogen atom, in a chain orin a ring, for example, which is sufficiently basic, such as anacid-addition salt with an inorganic acid, or “mineral acid”, such ashydrochloric, hydrobromic, hydroiodic, sulfuric, sulfamic, bisulfuric,phosphoric, or nitric acid, for example, or with an organic acid, suchas formic, acetic, acetoacetic, pyruvic, trifluoroacetic, propionic,butyric, hexanoic, heptanoic, undecanoic, lauric, benzoic, salicylic,2-(4-hydroxybenzoyl)-benzoic, camphoric, cinnamic,cyclopentanepropionic, digluconic, 3-hydroxy-2-naphthoic, nicotinic,pamoic, pectinic, 3-phenylpropionic, pivalic, 2-hydroxyethanesulfonic,itaconic, trifluoromethanesulfonic, dodecylsulfuric, ethanesulfonic,benzenesulfonic, para-toluenesulfonic, methanesulfonic,2-naphthalenesulfonic, naphthalinedisulfonic, camphorsulfonic acid,citric, tartaric, stearic, lactic, oxalic, malonic, succinic, malic,adipic, alginic, maleic, fumaric, D-gluconic, mandelic, ascorbic,glucoheptanoic, glycerophosphoric, aspartic, sulfosalicylic, orthiocyanic acid, for example.

Further, another suitably pharmaceutically acceptable salt of a compoundof the present invention which is sufficiently acidic, is an alkalimetal salt, for example a sodium or potassium salt, an alkaline earthmetal salt, for example a calcium, magnesium or strontium salt, or analuminium or a zinc salt, or an ammonium salt derived from ammonia orfrom an organic primary, secondary or tertiary amine having 1 to 20carbon atoms, such as ethylamine, diethylamine, triethylamine,ethyldiisopropylamine, monoethanolamine, diethanolamine,triethanolamine, dicyclohexylamine, dimethylaminoethanol,diethylaminoethanol, tris(hydroxymethyl)aminomethane, procaine,dibenzylamine, N-methylmorpholine, arginine, lysine,1,2-ethylenediamine, N-methylpiperidine, N-methyl-glucamine,N,N-dimethyl-glucamine, N-ethyl-glucamine, 1,6-hexanediamine,glucosamine, sarcosine, serinol, 2-amino-1,3-propanediol,3-amino-1,2-propanediol, 4-amino-1,2,3-butanetriol, or a salt with aquarternary ammonium ion having 1 to 20 carbon atoms, such astetramethylammonium, tetraethylammonium, tetra(n-propyl)ammonium,tetra(n-butyl)ammonium, N-benzyl-N,N,N-trimethylammonium, choline orbenzalkonium.

Those skilled in the art will further recognise that it is possible foracid addition salts of the claimed compounds to be prepared by reactionof the compounds with the appropriate inorganic or organic acid via anyof a number of known methods. Alternatively, alkali and alkaline earthmetal salts of acidic compounds of the present invention are prepared byreacting the compounds of the present invention with the appropriatebase via a variety of known methods.

The present invention includes all possible salts of the compounds ofthe present invention as single salts, or as any mixture of said salts,in any ratio.

In the present text, in particular in the Experimental Section, for thesynthesis of intermediates and of examples of the present invention,when a compound is mentioned as a salt form with the corresponding baseor acid, the exact stoichiometric composition of said salt form, asobtained by the respective preparation and/or purification process, is,in most cases, unknown.

Unless specified otherwise, suffixes to chemical names or structuralformulae relating to salts, such as “hydrochloride”, “trifluoroacetate”,“sodium salt”, or “x HCl”, “x CF₃COOH”, “x Na⁺”, for example, mean asalt form, the stoichiometry of which salt form not being specified.

This applies analogously to cases in which synthesis intermediates orexample compounds or salts thereof have been obtained, by thepreparation and/or purification processes described, as solvates, suchas hydrates, with (if defined) unknown stoichiometric composition.

Furthermore, the present invention includes all possible crystallineforms, or polymorphs, of the compounds of the present invention, eitheras single polymorph, or as a mixture of more than one polymorph, in anyratio.

In a distinct embodiment, the present invention relates to compounds offormula (I), supra, wherein

-   R¹ represents a group of the formula

-   -   in which    -   #¹ represents the point of attachment to the nitrogen atom,

-   R² represents a group selected from trifluoromethyl,    trifluoromethoxy, ethoxy, —C(═O)N(H)CH₃ and —S(═O)₂CH₃,

or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

In a preferred embodiment, the present invention relates to compoundsaccording to formula (I), supra, wherein

R¹ represents a group of the formula

-   -   in which    -   #¹ represents the point of attachment to the nitrogen atom,

R² represents a group selected from trifluoromethyl andtrifluoromethoxy.

or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

In accordance with a further preferred embodiment, the present inventioncovers compounds of general formula (I), supra, wherein

R¹ represents a (2S)-3,3,3-trifluoro-2-hydroxypropyl group of theformula

-   -   in which    -   #¹ represents the point of attachment to the nitrogen atom,

or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

In accordance with a further preferred embodiment, the present inventioncovers compounds of general formula (I), supra, wherein

R¹ represents a (2R)-3,3,3-trifluoro-2-hydroxypropyl group of theformula

-   -   in which    -   #¹ represents the point of attachment to the nitrogen atom,

or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

In accordance with a further preferred embodiment, the present inventioncovers compounds of general formula (I), supra, wherein

Ar represents a group of the formula

-   -   in which    -   #² represents the point of attachment to the nitrogen atom,

or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.

In a particular further embodiment of the first aspect, the presentinvention covers combinations of two or more of the above mentionedembodiments under the heading “further embodiments of the first aspectof the present invention”.

The present invention covers any sub-combination within any embodimentor aspect of the present invention of compounds of general formula (I),supra.

The present invention covers any sub-combination within any embodimentor aspect of the present invention of intermediate compounds of generalformula (II), (III), (IV), (V), (VI), and (VIII). The present inventioncovers the compounds of general formula (I) which are disclosed in theExample Section of this text, infra.

In accordance with a second aspect, the present invention covers methodsof preparing compounds of general formula (I) as defined supra, saidmethods comprising the step

[A] of allowing an intermediate compound of formula (II):

-   -   in which R¹ is as defined for the compound of general        formula (I) as defined supra, and    -   R³ represents a (C₁-C₄)-alkyl group, in particular a methyl        group,

to react in a first step in the presence of a base with a compound ofgeneral formula (III):

-   -   in which    -   R⁴ represents a (C₁-C₄)-alkyl group, in particular a methyl        group,

to give an intermediate compound, which is then allowed to react in thepresence of a base in a second step with a hydrazine compound of generalformula (IV) or a respective salt thereof

in which Ar is as defined for the compound of general formula (I) asdefined supra, thereby giving a compound of general formula (V):

-   -   in which R¹ and Ar are as defined for the compound of general        formula (I) as defined supra, and    -   R⁴ represents a (C₁-C₄)-alkyl group, in particular a methyl        group, followed by a subsequent step

[B] of allowing the compound of formula (V) obtained in step [A] toreact with ammonia thereby giving a compound of general formula (I):

-   -   in which R¹ and Ar are as defined for the compound of general        formula (I) as defined supra,

optionally followed by step

[C] conversion of the alcohols of general formula (I-A):

-   -   in which Ar is as defined for the compound of general        formula (I) as defined supra, to the ketones of general formula        (I-B):

-   -   in which Ar is as defined for the compound of general        formula (I) as defined supra, using known oxidation methods,

each [A], [B] and [C] optionally followed, where appropriate, by (i)separating the compounds of formula (I) thus obtained into theirrespective enantiomers, and/or (ii) converting the compounds of formula(I) into their respective hydrates, solvates, salts and/or hydrates orsolvates of the salts by treatment with the corresponding solventsand/or acids or bases.

The present invention covers methods of preparing compounds of thepresent invention of general formula (I), said methods comprising thesteps as described in the Experimental Section herein.

The schemes and procedures described below illustrate synthetic routesto the compounds of general formula (I) of the invention and are notintended to be limiting. It is clear to the person skilled in the artthat the order of transformations as exemplified in schemes 2, 3, 4, 5,6 and 7 can be modified in various ways. The order of transformationsexemplified in these schemes is therefore not intended to be limiting.In addition, interconversion of any of the substituents, R¹, R², R³, R⁴and Ar can be achieved before and/or after the exemplifiedtransformations. These modifications can be such as the introduction ofprotecting groups, cleavage of protecting groups, reduction or oxidationof functional groups, halogenation, metallation, substitution or otherreactions known to the person skilled in the art. These transformationsinclude those which introduce a functionality which allows for furtherinterconversion of substituents. Appropriate protecting groups and theirintroduction and cleavage are well-known to the person skilled in theart (see for example T. W. Greene and P. G. M. Wuts in Protective Groupsin Organic Synthesis, 3rd edition, Wiley 1999). Specific examples aredescribed in the subsequent paragraphs.

The multicomponent cyclization (II)→(V) is carried out by first reactingimidate of formula (II) with an acid chloride of formula (III) in thepresence of a base to form an intermediate which is in a subsequent stepreacted with the aryl hydrazine compound of formula (IV). Typically theformed intermediate is not isolated and the reaction over the two stepsis performed in one-pot. The arylhydrazine compound for formula (I) mayalso be used in form of its salts, such as a hydrochloride salt or atosylate salt. Under the alkaline reaction conditions, the hydrazinesalt will be reconverted into the free base form. The amount of baseadded may then be adjusted in this respect.

Suitable bases for both steps are typically tertiary amine bases, suchas N,N-diisopropylethylamine (DIPEA), triethylamine, triisopropylamine,N-methylimidazole, N-methylmorpholine, pyridine and4-(N,N-dimethylamino)pyridine. Preferably, N,N-diisopropylethylamine(DIPEA) is used as base. The reaction is performed in an inert organicsolvent, such as dichloromethane, 1,2-dichloroethane, methyl tert-butylether, tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, toluene,pyridine, ethyl acetate, acetonitrile or N,N-dimethylformamide, or in amixture of these solvents. Preferably tetrahydrofuran or dioxane or amixture thereof are used as solvents. The first step is generallycarried out at a temperature in the range of −10° C. to +120° C.,preferably at 0° C. The second step is generally carried out at atemperature in the range of +20° C. to +120° C., preferably at roomtemperature. Concomitant microwave irradiation may have a beneficialeffect in this reaction as well at a temperature in the range of +60° C.to +150° C., preferably at +120° C.

The aminolysis reaction (V)→(I) is usually carried out in a solution ofammonia. Suitable ammonia solutions for this step are saturated ammoniasolutions, in particular a solution of ammonia in methanol, ethanol,isopropanol, tetrahydrofuran, dioxane or water or a mixture thereof.

Preferably, a methanolic ammonia solution is used. The reaction ispreferably performed directly in the ammonia solution in the absence ofany further reaction solvent. This step is generally carried out at atemperature in the range of +20° C. to +120° C., preferably at roomtemperature. Concomitant microwave irradiation may have a beneficialeffect in this reaction as well at a temperature in the range of +60° C.to +150° C., preferably at +120° C.

The oxidation reaction (I-A)→(I-B) is carried out using customaryoxidation methods known from the literature [e.g. JOC, 1983, 48, 4155(Dess Martin oxidation); Tet Lett, 1994, 35, 3485 (IBX oxidation); JOC,1970, 35, 3589 (acid dichromate oxidation); Tet Lett, 1979, 399 (PDCoxidation); Tetrahedron, 1978, 34, 1651 (Swern oxidation)]. Thus, thealcohol group in the compounds of the general formula (I-A) ispreferably oxidized using Dess-Martin periodinane (DMP). In a typicalprocedure the reaction is carried out in dichloromethane at atemperature of 0° C. and subsequent warming up to room temperature.

Compounds of general formula (I) in which Ar comprises a sulfonesubstituent of the formula —S(═O)₂CH₃ may be prepared from therespective corresponding methylsulfanyl precursors by customaryoxidation methods known in the art, such as oxidation with3-chloroperoxybenzoic acid (mCPBA).

Compounds of general formula (II) as defined supra, can be prepared by amethod comprising the step

[a] of allowing an intermediate compound of formula (VI):

-   -   in which R¹ is as defined for the compound of general        formula (I) as defined sup ra, to react with a nitrile compound        of general formula (VII),

-   -   in which X represents a leaving group, such as chlorine,        bromine, iodine, mesylate or tosylate, in particular chlorine or        bromine,

thereby giving a compound of general formula (VIII)

-   -   in which R¹ is as defined for the compound of general        formula (I) as defined sup ra, followed by a subsequent step

[b] of allowing the compound of formula (VIII) obtained in step [a] toreact with a basic alcoholate, preferably sodium methanolate, therebygiving a compound of general formula (II),

-   -   in which R¹ is as defined for the compound of general        formula (I) as defined supra, and    -   R³ represents a (C₁-C₄)-alkyl group, in particular a methyl        group.

The N-alkylation reaction (VI)+(VII)→(VIII) (step [a]) is typicallycarried out in the presence of a base. Typical and exemplary basesinclude sodium carbonate, potassium carbonate, cesium carbonate,N,N-diisopropylethylamine, triethylamine, sodium tert-butylate orpotassium tert-butylate in acetonitrile, methylisobutylketone, dioxane,dimethylformamide, dimethylacetamide, N-methyl-pyrrolidinone,dimethylsulfoxide and sulfolane, preference is given to potassiumcarbonate in methylisobutylketone or acetonitrile. The reaction mayoptionally be carried out in an advantageous manner with addition of analkylation catalyst such as, for example, lithium bromide, sodiumiodide, lithium iodide, tetra-n-butylammoniumbromide,tetra-n-butylammoniumiodide or benzyltriethylammoniumchloride. Thereactions are generally carried out in a temperature range of from +40°C. to +120° C., preferably at from +60° C. to +80° C. The reactions canbe carried out at atmospheric, at elevated or at reduced pressure (forexample at from 0.5 to 5 bar); in general, the reactions are carried outat atmospheric pressure. It may be advantageous to slowly perform theaddition of the alkylation agent (VII) over a longer time span.

Conversion to the imidates of general formula (II) can be achieved viastandard reaction protocols known to the person skilled in the art (step[b]: (VIII)→(II)). The reaction is typically carried out under basicreactions conditions by reacting with a basic alcoholate. Typical bases,which may be used are sodium methanolate, sodium ethanolate, sodiumpropanolate, sodium isopropoxide, sodium tert-butylate or potassiumtert-butylate in methanol, ethanol, n-propanol, isopropanol, n-butanol,isobutanol and tert-butanol. Preference is given to sodium methanolatein methanol. The reactions are generally carried out in a temperaturerange of from +20 to +80° C., preferably at from +20 to +40° C.

Alternatively, the nitrile compounds of general formula (VIII) mayoptionally also be prepared as shown in the synthetic scheme 2 below:

The amide coupling (IX)→(X) can be carried out directly with the help ofa condensing agent or activating agent in the presence of a base or overtwo steps via an acyl chloride or carboxylic acid imidazolide. Typicalcondensation and activating agents for the amide formation in processsteps (IX)→(X) include, for example, carbodiimides such asN,N′-diethyl-, N, N′-dipropyl-, N,N′-diiso-propyl-,N,N′-dicyclohexylcarbodiimide (DCC) orN-(3-dimethylaminoisopropyl)-N′-ethylcarbo-diimide hydrochloride (EDC),phosgene derivatives such as N, N′-carbonyldiimidazole (CDI),1,2-oxazolium compounds such as2-ethyl-5-phenyl-1,2-oxazolium-3-sulphate or2-tert-butyl-5-methyl-isoxazolium perchlorate, acylamino compounds suchas 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, or isobutylchloroformate, propanephosphonic anhydride, diethyl cyano-phosphonate,bis(2-oxo-3-oxazolidinyl)phosphoryl chloride,benzotriazol-1-yloxytris(dimethyl-amino)phosphonium hexafluorophosphate,benzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate(PyBOP), O-(benzotriazol-1-yl)-N, N, N′,N′-tetramethyluroniumtetrafluoroborate (TBTU),O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate(HBTU), 2-(2-oxo-1-(2H)-pyridyl)-1,1,3,3-tetramethyluroniumtetrafluoroborate (TPTU),O-(7-aza-benzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate(HATU) or O-(1H-6-chloro-benzotriazol-1-yl)-1,1,3,3-tetramethyluroniumtetrafluoroborate (TCTU), optionally in combination with other additivessuch as 1-hydroxybenzotriazole (HOBt) or N-hydroxysuccinimide (HOSu).

The acyl chlorides are typically prepared with thionyl chloride oroxalyl chloride in an inert solvent like dichloromethane orN,N-dimethylformamide. It is also possible to use mixtures of thesolvents mentioned.

Typical and exemplary bases include sodium carbonate, potassiumcarbonate, cesium carbonate, N,N-diisopropylethylamine, triethylamine,sodium tert-butylate or potassium tert-butylate in acetonitrile,methylisobutylketone, dioxane, dimethylformamide, dimethylacetamide,N-methyl-pyrrolidinone, dimethylsulfoxide and sulfolane, preference isgiven to potassium carbonate in methylisobutylketone or acetonitrile.The reaction may optionally be carried out in an advantageous mannerwith addition of an alkylation catalyst such as, for example, lithiumbromide, sodium iodide, lithium iodide, tetra-n-butylammoniumbromide,tetra-n-butylammoniumiodide or benzyltriethylammoniumchloride. Theconversion to the nitrile (X)→(XI) can be carried out with the help ofdehydrating agent. Typical dehydrating agents include, for exampletrifluoroacetic acid anhydride, phosphorous pentoxide (P₄O₁₀),phosphoryl chloride (POCl₃), phosphorous penta-chloride (PCl₅),CCl₄—PPh₃ (Appel reagent), hexamethylphosphoramide (HMPA); methylN-(triethylammoniumsulfonyl)carbamate (Burgess reagent),(Chloromethylene)dimethyliminium chloride (Vilsmeier reagent), oxalylchloride/DMSO and thionylchloride (SOCl₂).

Typical and exemplary solvents for both steps (IX)→(X) and (X)→(XI)include for example, ethers such as diethyl ether, dioxane,tetrahydrofuran, glycol dimethyl ether or diethylene glycol dimethylether, hydrocarbons such as benzene, toluene, xylene, hexane,cyclohexane or mineral oil, fractions, halogenated hydrocarbons such asdichloromethane, trichloromethane, carbon tetrachloride,1,2-dichloroethane, trichloroethylene or chlorobenzene, or othersolvents such as acetone, ethyl acetate, acetonitrile, pyridine,dimethyl sulphoxide, N,N-dimethylformamide, N,N′-dimethylpropyleneurea(DMPU) or N-methylpyrrolidone (NMP). It is also possible to use mixturesof the solvents mentioned.

In a typical and preferred procedure the carboxylic acid (IX) is firstreacted with pivaloyl chloride in the presence of pyridine to form anintermediate which is in a subsequent step reacted with ammonia.Typically the formed intermediate is not isolated and the reaction overthe two steps is performed in one-pot. Suitable as bases for the firststep are preferably, pyridine, 4-(N,N-dimethyl-amino)pyridine orN,N-diisopropylethylamine (DIPEA). The conversion of carboxamide (X)into nitrile (VIII) is then typically performed by reaction withtrifluoroacetic anhydride. Both reactions are conducted in an inertorganic solvent, preferably tetrahydrofuran.

The compounds of formula (VI) and (IX) can be synthesized by theprocedures described in Int. Pat. Appl. WO 2010/105770 and WO2011/104322 (see also synthesis schemes 3 and 4 below).

The compounds of the formulae (III), (IV) and (VII) are eithercommercially available, known from the literature, or can be preparedfrom readily available starting materials by adaptation of standardmethods described in the literature. Detailed procedures and literaturereferences for preparing the starting materials can also be found in theExperimental Part in the section on the preparation of the startingmaterials and intermediates.

The preparation of the compounds of the invention may be illustrated bymeans of the following synthesis schemes:

The compounds of general formula (I) of the present invention can beconverted to any salt, preferably pharmaceutically acceptable salts, asdescribed herein, by any method which is known to the person skilled inthe art. Similarly, any salt of a compound of general formula (I) of thepresent invention can be converted into the free compound, by any methodwhich is known to the person skilled in the art.

The compounds of the present invention have valuable pharmacologicalproperties and can be used for the prevention and/or treatment ofvarious diseases and disease-induced states in humans and other mammals.Compounds of general formula (I) of the present invention demonstrate avaluable pharmacological spectrum of action and pharmacokinetic profile,both of which could not have been predicted. Compounds of the presentinvention have surprisingly been found to effectively inhibit thevasopressin V1a receptor and it is possible therefore that saidcompounds be used for the treatment and/or prevention of diseases,preferably renal and cardiovascular diseases in humans and animals.

In the context of the present invention, the term “treatment” or“treating” includes inhibiting, delaying, relieving, mitigating,arresting, reducing, or causing the regression of a disease, disorder,condition, or state, the development and/or progression thereof, and/orthe symptoms thereof. The term “prevention” or “preventing” includesreducing the risk of having, contracting, or experien-cing a disease,disorder, condition, or state, the development and/or progressionthereof, and/or the symptoms thereof. The term prevention includesprophylaxis. Treatment or prevention of a disorder, disease, condition,or state may be partial or complete.

Throughout this document, for the sake of simplicity, the use ofsingular language is given preference over plural language, but isgenerally meant to include the plural language if not otherwise stated.For example, the expression “A method of treating a disease in apatient, comprising administering to a patient an effective amount of acompound of formula (I)” is meant to include the simultaneous treatmentof more than one disease as well as the administration of more than onecompound of formula (I).

The compounds of the present invention are highly potent and inparticular selective antagonists of the vasopressin V1a receptor. Thecompounds of the invention are therefore expected to be highly valuableas therapeutic agents for the treatment and/or prevention of diseases,in particular for the treatment and/or prevention of renal andcardiovascular diseases.

As used herein, the term “vasopressin V1a receptor antagonist” refers toa compound that functions by inhibiting (partially or completely) orblocking the vasopressin V1a receptor, thereby preventing activation ofthe receptor by vasopressin.

In one embodiment, compounds described herein are active at the V1areceptor. In another embodiment compounds described herein exhibitinhibition of the V1a receptor according to the study in B-1 with anIC₅₀<20 nM. In another embodiment compounds described herein exhibitinhibition of the V1a receptor according to the study in B-1 with anIC₅₀<10 nM. In another embodiment compounds described herein exhibitinhibition of the V1a receptor according to the study in B-1 with anIC₅₀<5 nM. In another embodiment compounds described herein exhibitinhibition of the V1a receptor according to the study in B-1 with anIC₅₀<1 nM.

In a further embodiment, compounds described herein are selectivelyactive at the V1a receptor, and are less active, substantially lessactive, and/or inactive at other vasopressin receptors, such as the V1band/or V2 subtypes. In another embodiment, compounds described hereinare at least 10-fold selective for the V1a receptor compared to the V2receptor as determined according to the study in B-1. In anotherembodiment, compounds described herein are at least 15-fold selectivefor the V1a receptor compared to the V2 receptor as determined accordingto the study in B-1. In another embodiment, compounds described hereinare at least 20-fold selective for the V1a receptor compared to the V2receptor as determined according to the study in B-1. In anotherembodiment, compounds described herein are at least 30-fold selectivefor the V1a receptor compared to the V2 receptor as determined accordingto the study in B-1. In another embodiment, compounds described hereinare at least 40-fold selective for the V1a receptor compared to the V2receptor as determined according to the study in B-1.

The compounds according to the invention are suitable for the treatmentand/or prevention of renal diseases, in particular of acute and chronickidney diseases, diabetic kidney diseases, and of acute and chronicrenal failure. The general terms ‘renal disease’ or ‘kidney disease’describe a class of conditions in which the kidneys fail to filter andremove waste products from the blood. There are two major forms ofkidney disease: acute kidney disease (acute kidney injury, AKI) andchronic kidney disease (CKD). The compounds according to the inventionmay further be used for the treatment and/or prevention of sequelae ofacute kidney injury arising from multiple insults such asischemia-reperfusion injury, radiocontrast administration,cardiopulmonary bypass surgery, shock and sepsis. In the sense of thepresent invention, the term renal failure or renal insufficiencycomprises both acute and chronic manifestations of renal insufficiency,as well as underlying or related kidney diseases such as renalhypoperfusion, intradialytic hypotension, obstructive uropathy,glomerulopathies, IgA nephropathy, glomerulonephritis, acuteglomerulonephritis, glomerulosclerosis, tubulointerstitial diseases,nephropathic diseases such as primary and congenital kidney disease,nephritis, Alport syndrome, kidney inflammation, immunological kidneydiseases such as kidney transplant rejection, immune complex-inducedkidney diseases, nephropathy induced by toxic substances, contrastmedium-induced nephropathy; minimal change glomerulonephritis (lipoid);Membranous glomerulonephritis; focal segmental glomerulosclerosis(FSGS); hemolytic uremic syndrome (HUS), amyloidosis, Goodpasture'ssyndrome, Wegener's granulomatosis, Purpura Schonlein-Henoch, diabeticand non-diabetic nephropathy, pyelonephritis, renal cysts,nephrosclerosis, hypertensive nephrosclerosis and nephrotic syndrome,which can be characterized diagnostically, for example, by abnormallyreduced creatinine and/or water excretion, abnormally increased bloodconcentrations of urea, nitrogen, potassium and/or creatinine, alteredactivity of renal enzymes such as, for example, glutamyl synthetase,altered urine osmolarity or urine volume, increased microalbuminuria,macroalbuminuria, lesions of glomeruli and arterioles, tubulardilatation, hyperphosphataemia and/or the need for dialysis. The presentinvention also comprises the use of the compounds according to theinvention for the treatment and/or prevention of sequelae of renalinsufficiency, for example pulmonary edema, heart failure, uraemia,anaemia, electrolyte disturbances (e.g. hyperkalaemia, hyponatraemia)and disturbances in bone and carbohydrate metabolism. The compoundsaccording to the invention are also suitable for the treatment and/orprevention of polycystic kidney disease (PCKD) and of the syndrome ofinadequate ADH secretion (SIADH).

Cardiovascular diseases in this context that may be treated and/orprevented with the compounds of the invention include, but are notlimited to, the following: acute and chronic heart failure includingworsening chronic heart failure (or hospitalization for heart failure)and including congestive heart failure, arterial hypertension, resistanthypertension, arterial pulmonary hypertension, coronary heart disease,stable and unstable angina pectoris, atrial and ventricular arrhythmias,disturbances of atrial and ventricular rhythm and conductiondisturbances, for example atrioventricular blocks of degree I-III (AVBI-III), supraventricular tachyarrhythmia, atrial fibrillation, atrialflutter, ventricular fibrillation, ventricular flutter, ventriculartachyarrhythmia, torsade-de-pointes tachycardia, atrial and ventricularextrasystoles, AV-junction extrasystoles, sick-sinus syndrome, syncopes,AV-node re-entry tachycardia and Wolff-Parkinson-White syndrome, acutecoronary syndrome (ACS), autoimmune heart diseases (pericarditis,endocarditis, valvulitis, aortitis, cardiomyopathies), shock such ascardiogenic shock, septic shock and anaphylactic shock, aneurysms, Boxercardiomyopathy (premature ventricular contraction), furthermorethromboembolic diseases and ischaemias such as peripheral perfusiondisturbances, reperfusion injury, arterial and venous thromboses,myocardial insufficiency, endothelial dysfunction, micro- andmacrovascular damage (vasculitis) and for preventing restenoses such asafter thrombolysis therapies, percutaneous transluminal angioplasty(PTA), percutaneous transluminal coronary angioplasty (PTCA), hearttransplantation and bypass operations, arteriosclerosis, disturbances oflipid metabolism, hypolipoproteinaemias, dyslipidemias,hypertriglyceridemias, hyperlipidemias and combined hyperlipidemias,hyper-cholesterolaemias, abetalipoproteinaemia, sitosterolemia,xanthomatosis, Tangier disease, adipositas, obesity, metabolic syndrome,transitory and ischemic attacks, stroke, inflammatory cardiovasculardiseases, peripheral and cardiac vascular diseases, peripheralcirculation disorders, spasms of the coronary arteries and peripheralarteries, and edema such as, for example, pulmonary edema, cerebraledema, renal edema and heart failure-related edema.

In the sense of the present invention, the term heart failure alsoincludes more specific or related disease forms such as right heartfailure, left heart failure, global insufficiency, ischemiccardiomyopathy, dilatative cardiomyopathy, congenital heart defects,heart valve defects, heart failure with heart valve defects, mitralvalve stenosis, mitral valve insufficiency, aortic valve stenosis,aortic valve insufficiency, tricuspidal stenosis, tricuspidalinsufficiency, pulmonary valve stenosis, pulmonary valve insufficiency,combined heart valve defects, heart muscle inflammation (myocarditis),chronic myocarditis, acute myocarditis, viral myocarditis, diabeticheart failure, alcohol-toxic cardiomyopathy, cardiac storage diseases,heart failure with preserved ejection fraction (HFpEF or diastolic heartfailure), and heart failure with reduced ejection fraction (HFrEF orsystolic heart failure).

The compounds of the present invention may be particularly useful forthe treatment and/or prevention of the cardiorenal syndrome (CRS) andits various subtypes. This term embraces certain disorders of the heartand kidneys whereby acute or chronic dysfunction in one organ may induceacute or chronic dysfunction of the other.

Moreover, the compounds according to the invention may be used for thetreatment and/or prevention of peripheral arterial disease (PAD)including claudication and including critical limb ischemia, coronarymicrovascular dysfunction (CMD) including CMD type 1-4, primary andsecondary Raynaud's phenomenon, microcirculation disturbances,peripheral and autonomic neuropathies, diabetic microangiopathies,diabetic retinopathy, diabetic limb ulcers, gangrene, CREST syndrome,erythematous disorders, rheumatic diseases and for promoting woundhealing.

Furthermore, the compounds of the invention are suitable for treatingurological diseases and diseases of the male and female urogenitalsystem such as, for example, benign prostatic syndrome (BPS), benignprostatic hyperplasia (BPH), benign prostatic enlargement (BPE), bladderoutlet obstruction (BOO), lower urinary tract syndromes (LUTS),neurogenic overactive bladder (OAB), interstitial cystitis (IC), urinaryincontinence (UI) such as, for example, mixed, urge, stress and overflowincontinence (MUI, UUI, SUI, OUI), pelvic pains, erectile dysfunction,dysmenorrhea and endometriosis.

The compounds according to the invention may also be used for thetreatment and/or prevention of inflammatory diseases, asthmaticdiseases, chronic obstructive pulmonary disease (COPD), acuterespiratory distress syndrome (ARDS), acute lung injury (ALI),alpha-1-antitrypsin deficiency (AATD), pulmonary fibrosis, pulmonaryemphysema (e.g. smoking-induced pulmonary emphysema) and cystic fibrosis(CF). In addition, the compounds of the invention may be used for thetreatment and/or prevention of pulmonary arterial hypertension (PAH) andother forms of pulmonary hypertension (PH) including pulmonaryhypertension associated with left ventricular disease, HIV infection,sickle cell anaemia, thromboembolism (CTEPH), sarcoidosis, chronicobstructive pulmonary disease (COPD) or pulmonary fibrosis.

Additionally, the compounds according to the invention may be used forthe treatment and/or prevention of liver cirrhosis, ascites, diabetesmellitus and diabetic complications such as, for example, neuropathy andnephropathy.

Further, the compounds of the invention are suitable for the treatmentand/or prevention of central nervous disorders such as anxiety states,depression, glaucoma, cancer such as particular pulmonary tumors, andcircadian rhythm misalignment such as jet lag and shift work.

Furthermore, the compounds according to the invention may be useful forthe treatment and/or prevention of pain conditions, diseases of theadrenals such as, for example, pheochromocytoma and adrenal apoplexy,diseases of the intestine such as, for example, Crohn's disease anddiarrhea, menstrual disorders such as, for example, dysmenorrhea,endometriosis, preterm labor and tocolysis.

Due to their activity and selectivity profile, the compounds of thepresent invention are believed to be particularly suitable for thetreatment and/or prevention of acute and chronic kidney diseasesincluding diabetic nephropathy, acute and chronic heart failure,preeclampsia, peripheral arterial disease (PAD), coronary microvasculardysfunction (CMD), Raynaud's syndrome and dysmenorrhea.

The diseases mentioned above have been well characterized in humans, butalso exist with a com-parable etiology in other mammals, and may betreated in those with the compounds and methods of the presentinvention.

Thus, the present invention further relates to the use of the compoundsaccording to the invention for the treatment and/or prevention ofdiseases, especially of the aforementioned diseases. The presentinvention further relates to the use of the compounds according to theinvention for preparing a pharmaceutical composition for the treatmentand/or prevention of diseases, especially of the aforementioneddiseases.

The present invention further relates to the use of the compoundsaccording to the invention in a method for the treatment and/orprevention of diseases, especially of the aforementioned diseases.

The present invention further relates to a method for the treatmentand/or prevention of diseases, especially of the aforementioneddiseases, by using an effective amount of at least one of the compoundsaccording to the invention.

In accordance with another aspect, the present invention coverspharmaceutical combinations, in particular medicaments, comprising atleast one compound of general formula (I) of the present invention andat least one or more further active ingredients, in particular for thetreatment and/or prevention of diseases, especially of theaforementioned diseases.

Particularly, the present invention covers a pharmaceutical combination,which comprises:

-   -   one or more first active ingredients, in particular compounds of        general formula (I) as defined supra, and    -   one or more further active ingredients, in particular for the        treatment and/or prevention of diseases, especially of the        aforementioned diseases.

The term “combination” in the present invention is used as known topersons skilled in the art, it being possible for said combination to bea fixed combination, a non-fixed combination or a kit-of-parts.

A “fixed combination” in the present invention is used as known topersons skilled in the art and is defined as a combination wherein, forexample, a first active ingredient, such as one or more compounds ofgeneral formula (I) of the present invention, and a further activeingredient are present together in one unit dosage or in one singleentity. One example of a “fixed combination” is a pharmaceuticalcomposition wherein a first active ingredient and a further activeingredient are present in admixture for simultaneous administration,such as in a formulation. Another example of a “fixed combination” is apharmaceutical combination wherein a first active ingredient and afurther active ingredient are present in one unit without being inadmixture.

A non-fixed combination or “kit-of-parts” in the present invention isused as known to persons skilled in the art and is defined as acombination wherein a first active ingredient and a further activeingredient are present in more than one unit. One example of a non-fixedcombination or kit-of-parts is a combination wherein the first activeingredient and the further active ingredient are present separately. Itis possible for the components of the non-fixed combination orkit-of-parts to be administered separately, sequentially,simultaneously, concurrently or chronologically staggered.

The compounds of the present invention can be administered as the solepharmaceutical agent or in combination with one or more otherpharmaceutically active ingredients where the combination causes nounacceptable adverse effects. The present invention also covers suchpharmaceutical combinations. For example, the compounds of the presentinvention can be combined with known agents for the treatment and/orprevention of diseases, especially of the aforementioned diseases.

In particular, the compounds of the present invention may be used infixed or separate combination with

-   -   antithrombotic agents, for example and preferably from the group        of platelet aggregation inhibitors, anticoagulants and        profibrinolytic substances;    -   blood pressure lowering agents, for example and preferably from        the group of calcium antagonists, angiotensin AII antagonists,        ACE inhibitors, NEP inhibitors, vasopeptidase inhibitors,        endothelin antagonists, renin inhibitors, alpha-blockers,        beta-blockers, mineralocorticoid receptor antagonists and        diuretics;    -   antidiabetic agents (hypoglycemic or antihyperglycemic agents),        such as for example and preferably insulin and derivatives,        sulfonylureas, biguanides, thiazolidinediones, acarbose, DPP4        inhibitors, GLP-1 analogues, or SGLT inhibitors (gliflozins).    -   organic nitrates and NO-donors, for example sodium        nitroprusside, nitroglycerin, isosorbide mononitrate, isosorbide        dinitrate, molsidomine or SIN-1, and inhalational NO;    -   compounds that inhibit the degradation of cyclic guanosine        monophosphate (cGMP), for example inhibitors of        phosphodiesterases (PDE) 1, 2, 5 and/or 9, in particular PDE-5        inhibitors such as sildenafil, vardenafil, tadalafil, udenafil,        dasantafil, avanafil, mirodenafil, lodenafil, CTP-499 or        PF-00489791;    -   natriuretic peptides, such as for example atrial natriuretic        peptide (ANP, anaritide), B-type natriuretic peptide or brain        natriuretic peptide (BNP, nesiritide), C-type natriuretic        peptide (CNP) or urodilatin;    -   calcium sensitizers, such as for example and preferably        levosimendan;    -   NO- and heme-independent activators of soluble guanylate cyclase        (sGC), for example and with preference the compounds described        in WO 01/19355, WO 01/19776, WO 01/19778, WO 01/19780, WO        02/070462 and WO 02/070510;    -   NO-independent, but heme-dependent stimulators of guanylate        cyclase (sGC), for example and with preference the compounds        described in WO 00/06568, WO 00/06569, WO 02/42301, WO        03/095451, WO 2011/147809, WO 2012/004258, WO 2012/028647 and WO        2012/059549;    -   agents, that stimulates the synthesis of cGMP, for example and        with preference sGC modulators, for example and with preference        riociguat, cinaciguat, vericiguat or BAY 1101042;    -   inhibitors of human neutrophil elastase (HNE), such as for        example sivelestat or DX-890 (reltran);    -   compounds inhibiting the signal transduction cascade, in        particular tyrosine and/or serine/threo-nine kinase inhibitors,        such as for example nintedanib, dasatinib, nilotinib, bosutinib,        regora-fenib, sorafenib, sunitinib, cediranib, axitinib,        telatinib, imatinib, brivanib, pazopanib, vatalanib, gefitinib,        erlotinib, lapatinib, canertinib, lestaurtinib, pelitinib,        semaxanib or tandutinib;    -   compounds influencing the energy metabolism of the heart, such        as for example and preferably etomoxir, dichloroacetate,        ranolazine or trimetazidine, or full or partial adenosine A1        receptor agonists as GS-9667 (previously known as CVT-3619),        capadenoson and neladenoson bialanate (BAY 1067197);    -   compounds influencing the heart rate, such as for example and        preferably ivabradine;    -   cardiac myosin activators, such as for example and preferably        omecamtiv mecarbil (CK-1827452);    -   anti-inflammatory drugs such as non-steroidal anti-inflammatory        drugs (NSAIDs) including acetylsalicylic acid (aspirin),        ibuprofen and naproxen, glucocorticoids such as for example and        preferably prednison, prednisolon, methylprednisolon,        triamcinolon, dexamethason, beclomethason, betamethason,        flunisolid, budesonid or fluticason, 5-aminosalicylic acid        derivatives, leukotriene antagonists, TNF-alpha inhibitors and        chemokine receptor antagonists such as CCR1, 2 and/or 5        inhibitors;    -   fat metabolism altering agents, for example and preferably from        the group of thyroid receptor agonists, cholesterol synthesis        inhibitors, such as for example and preferably HMG-CoA-reductase        or squalene synthesis inhibitors, ACAT inhibitors, CETP        inhibitors, MTP inhibitors, PPAR-alpha, PPAR-gamma and/or        PPAR-delta agonists, cholesterol absorption inhibitors, lipase        inhibitors, polymeric bile acid adsorbers, bile acid        reabsorption inhibitors and lipoprotein(a) antagonists.

Antithrombotic agents are preferably to be understood as compounds fromthe group of platelet aggregation inhibitors, anticoagulants andprofibrinolytic substances.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a plateletaggregation inhibitor, for example and preferably aspirin, clopidogrel,ticlopidine or dipyridamole.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a thrombin inhibitor,for example and preferably ximelagatran, dabigatran, melagatran,bivalirudin or enoxaparin.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a GPIIb/IIIaantagonist, for example and preferably tirofiban or abcixi-mab.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a factor Xainhibitor, for example and preferably rivaroxaban, apixaban, otamixaban,fidexaban, razaxaban, fondaparinux, idraparinux, DU-176b, PMD-3112,YM-150, KFA-1982, EMD-503982, MCM-17, MLN-1021, DX 9065a, DPC 906, JTV803, SSR-126512 or SSR-128428.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with heparin or a lowmolecular weight (LMW) heparin derivative.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a vitamin Kantagonist, for example and preferably coumarin.

Blood pressure lowering agents are preferably to be understood ascompounds from the group of calcium antagonists, angiotensin AIIantagonists, ACE inhibitors, NEP inhibitors, vasopeptidase inhibitors,endothelin antagonists, renin inhibitors, alpha-blockers, beta-blockers,mineralocorticoid receptor antagonists and diuretics.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a calcium antagonist,for example and preferably nifedipine, amlodipine, verapamil ordiltiazem.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an alpha-1-receptorblocker, for example and preferably prazosin or tamsulosin.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a beta-blocker, forexample and preferably propranolol, atenolol, timolol, pindolol,alprenolol, oxprenolol, penbutolol, bupranolol, metipranolol, nadolol,mepindolol, carazolol, sotalol, metoprolol, betaxolol, celiprolol,bisoprolol, carteolol, esmolol, labetalol, carvedilol, adaprolol,landiolol, nebivolol, epanolol or bucindolol.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an angiotensin AIIreceptor antagonist, for example and preferably losartan, candesartan,valsartan, telmisartan, irbesartan, olmesartan, eprosartan, embursartanor azilsartan.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a vasopeptidaseinhibitor or inhibitor of neutral endopeptidase (NEP), such as forexample and preferably sacubitril, omapatrilat or AVE-7688.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a dual angiotensinAII receptor antagonist/NEP inhibitor (ARNI), for example and preferablyLCZ696.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an ACE inhibitor, forexample and preferably enalapril, captopril, lisinopril, ramipril,delapril, fosinopril, quinopril, perindopril, benazepril or trandopril.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an endothelinantagonist, for example and preferably bosentan, darusentan,ambrisentan, tezosentan, sitaxsentan, avosentan, macitentan oratrasentan.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a renin inhibitor,for example and preferably aliskiren, SPP-600 or SPP-800.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a mineralocorticoidreceptor antagonist, for example and preferably finerenone,spironolactone, canrenone, potassium canrenoate, eplerenone, esaxerenone(CS-3150), or apararenone (MT-3995).

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a diuretic, such asfor example and preferably furosemide, bumetanide, piretanide,torsemide, bendroflumethiazide, chlorothiazide, hydrochlorothiazide,xipamide, indapamide, hydroflumethiazide, methyclothiazide,polythiazide, trichloromethiazide, chlorothalidone, metolazone,quinethazone, acetazolamide, dichlorophenamide, methazolamide,glycerine, isosorbide, mannitol, amiloride or triamterene.

Fat metabolism altering agents are preferably to be understood ascompounds from the group of CETP inhibitors, thyroid receptor agonists,cholesterol synthesis inhibitors such as HMG-CoA-reductase or squalenesynthesis inhibitors, ACAT inhibitors, MTP inhibitors, PPAR-alpha,PPAR-gamma and/or PPAR-delta agonists, cholesterol absorptioninhibitors, polymeric bile acid adsorbers, bile acid reabsorptioninhibitors, lipase inhibitors and lipoprotein(a) antagonists.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a CETP inhibitor, forexample and preferably dalcetrapib, anacetrapib, BAY 60-5521 orCETP-vaccine (Avant).

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a thyroid receptoragonist, for example and preferably D-thyroxin, 3,5,3′-triiodothyronin(T3), CGS 23425 or axitirome (CGS 26214).

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an HMG-CoA-reductaseinhibitor from the class of statins, for example and preferablylovastatin, simvastatin, pravastatin, fluvastatin, atorvastatin,rosuvastatin or pitavastatin.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a squalene synthesisinhibitor, for example and preferably BMS-188494 or TAK-475.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an ACAT inhibitor,for example and preferably avasimibe, melinamide, pactimibe, eflucimibeor SMP-797.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an MTP inhibitor, forexample and preferably implitapide, R-103757, BMS-201038 or JTT-130.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a PPAR-gamma agonist,for example and preferably pioglitazone or rosiglitazone.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a PPAR-delta agonist,for example and preferably GW 501516 or BAY 68-5042.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a cholesterolabsorption inhibitor, for example and preferably ezetimibe, tiqueside orpamaqueside.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a lipase inhibitor,for example and preferably orlistat.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a polymeric bile acidadsorber, for example and preferably cholestyr-amine, colestipol,colesolvam, CholestaGel or colestimide.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a bile acidreabsorption inhibitor, for example and preferably ASBT (=IBAT)inhibitors such as AZD-7806, S-8921, AK-105, BARI-1741, SC-435 orSC-635.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a lipoprotein(a)antagonist, for example and preferably gemcabene calcium (CI-1027) ornicotinic acid.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a TGF betaantagonist, by way of example and with preference pirfenidone orfresolimumab.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with HIF-PH inhibitors, byway of example and with preference molidustat or roxadustat.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a CCR2 antagonist, byway of example and with preference CCX-140.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a TNFalphaantagonist, by way of example and with preference adalimumab.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a galectin-3inhibitor, by way of example and with preference GCS-100.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a BMP-7 agonist, byway of example and with preference THR-184.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a NOX1/4 inhibitor,by way of example and with preference GKT-137831.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a medicament whichaffects the vitamin D metabolism, by way of example and with preferencecholecalciferol or paracalcitol.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a cytostatic agent,by way of example and with preference cyclophosphamide.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an immunosuppressiveagent, by way of example and with preference ciclosporin.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a phosphate binder,by way of example and with preference sevelamer or lanthanum carbonate.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a calcimimetic fortherapy of hyperparathyroidism.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with agents for irondeficit therapy, by way of example and with preference iron products.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with agents for thetherapy of hyperurikaemia, by way of example and with preferenceallopurinol or rasburicase.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with glycoprotein hormonefor the therapy of anaemia, by way of example and with preferenceerythropoietin.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with biologics for immunetherapy, by way of example and with preference abatacept, rituximab,eculizumab or belimumab.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with Jak inhibitors, byway of example and with preference ruxolitinib, tofacitinib,baricitinib, CYT387, GSK2586184, lestaurtinib, pacritinib (SB1518) orTG101348.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with prostacyclin analogsfor therapy of microthrombi.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an alkali therapy, byway of example and with preference sodium bicarbonate.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an mTOR inhibitor, byway of example and with preference everolimus or rapamycin.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an NHE3 inhibitor, byway of example and with preference AZD1722.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with an eNOS modulator, byway of example and with preference sapropterin.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with a CTGF inhibitor, byway of example and with preference FG-3019.

In a preferred embodiment of the invention, the compounds according tothe invention are administered in combination with antidiabetics(hypoglycemic or antihyperglycemic agents), such as for example andpreferably insulin and derivatives, sulfonylureas such as tolbutamide,carbutamide, acetohexamide, chlorpropamide, glipizide, gliclazide,glibenclamide, glyburide, glibornuride, gliquidone, glisoxepide,glyclopyramide, glimepiride, JB253 and JB558, meglitinides such asrepaglinide and nateglinide, biguanides such as metformin and buformin,thiazolidinediones such as rosiglitazone and pioglitazone,alpha-glucosidase inhibitors such as miglitol, acarbose and voglibose,DPP4 inhibitors such as vildagliptin, sitagliptin, saxagliptin,linagliptin, alogliptin, septagliptin and teneligliptin, GLP-1 analoguessuch as exenatide (also exendin-4, liraglutide, lixisenatide andtaspoglutide, or SGLT inhibitors (gliflozins) such as canagliflozin,dapagliflozin and empagliflozin.

In a particularly preferred embodiment, the compounds of the presentinvention are administered in combination with one or more additionaltherapeutic agents selected from the group consisting of diuretics,angiotensin AII antagonists, ACE inhibitors, beta-receptor blockers,mineralocorticoid receptor antagonists, antidiabetics, organic nitratesand NO donors, activators and stimulators of the soluble guanylatecyclase (sGC), and positive-inotropic agents.

In a further particularly preferred embodiment, the compounds of thepresent invention are administered in combination with one or moreadditional therapeutic agents selected from the group consisting ofdiuretics, angiotensin AII antagonists, ACE inhibitors, beta-receptorblockers, mineralocorticoid receptor antagonists, antidiabetics, organicnitrates and NO donors, activators and stimulators of the solubleguanylate cyclase (sGC), antiinflammatory agents, immunosuppressiveagents, phosphate binders and/or compounds which modulate vitamin Dmetabolism.

Thus, in a further embodiment, the present invention relates topharmaceutical compositions comprising at least one of the compoundsaccording to the invention and one or more additional therapeutic agentsfor the treatment and/or prevention of diseases, especially of theaforementioned diseases.

Furthermore, the compounds of the present invention may be utilized, assuch or in compositions, in research and diagnostics, or as analyticalreference standards and the like, which are well known in the art.

When the compounds of the present invention are administered aspharmaceuticals, to humans and other mammals, they can be given per seor as a pharmaceutical composition containing, for example, 0.1% to99.5% (more preferably, 0.5% to 90%) of active ingredient in combinationwith one or more pharmaceutically acceptable excipients.

Thus, in another aspect, the present invention relates to pharmaceuticalcompositions comprising at least one of the compounds according to theinvention, conventionally together with one or more inert, non-toxic,pharmaceutically acceptable excipients, and to the use thereof for thetreatment and/or prevention of diseases, especially of theaforementioned diseases.

It is possible for the compounds according to the invention to havesystemic and/or local activity. For this purpose, they can beadministered in a suitable manner, such as, for example, via the oral,parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal,vaginal, dermal, transdermal, conjunctival, otic route or as an implantor stent.

For these administration routes, it is possible for the compoundsaccording to the invention to be administered in suitable administrationforms.

For oral administration, it is possible to formulate the compoundsaccording to the invention to dosage forms known in the art that deliverthe compounds of the invention rapidly and/or in a modified manner, suchas, for example, tablets (uncoated or coated tablets, for example withenteric or controlled release coatings that dissolve with a delay or areinsoluble), orally-disintegrating tablets, films/wafers,films/lyophylisates, capsules (for example hard or soft gelatinecapsules), sugar-coated tablets, granules, pellets, powders, emulsions,suspensions, aerosols or solutions. It is possible to incorporate thecompounds according to the invention in crystalline and/or amorphisedand/or dissolved form into said dosage forms.

Parenteral administration can be effected with avoidance of anabsorption step (for example intravenous, intraarterial, intracardial,intraspinal or intralumbal) or with inclusion of absorption (for exampleintramuscular, subcutaneous, intracutaneous, percutaneous orintraperitoneal). Administration forms which are suitable for parenteraladministration are, inter alia, preparations for injection and infusionin the form of solutions, suspensions, emulsions, lyophylisates orsterile powders.

Examples which are suitable for other administration routes arepharmaceutical forms for inhalation [inter alia powder inhalers,nebulizers], nasal drops, nasal solutions, nasal sprays;tablets/films/wafers/capsules for lingual, sublingual or buccaladministration; suppositories; eye drops, eye ointments, eye baths,ocular inserts, ear drops, ear sprays, ear powders, ear-rinses, eartampons; vaginal capsules, aqueous suspensions (lotions, mixturaeagitandae), lipophilic suspensions, emulsions, ointments, creams,transdermal therapeutic systems (such as, for example, patches), milk,pastes, foams, dusting powders, implants or stents.

The compounds according to the invention can be incorporated into thestated administration forms. This can be effected in a manner known perse by mixing with pharmaceutically suitable excipients. Pharmaceuticallysuitable excipients include, inter alia,

-   -   fillers and carriers (for example cellulose, microcrystalline        cellulose (such as, for example, Avicel®), lactose, mannitol,        starch, calcium phosphate (such as, for example, Di-Cafos®)),    -   ointment bases (for example petroleum jelly, paraffins,        triglycerides, waxes, wool wax, wool wax alcohols, lanolin,        hydrophilic ointment, polyethylene glycols),    -   bases for suppositories (for example polyethylene glycols, cacao        butter, hard fat),    -   solvents (for example water, ethanol, isopropanol, glycerol,        propylene glycol, medium chain-length triglycerides fatty oils,        liquid polyethylene glycols, paraffins),    -   surfactants, emulsifiers, dispersants or wetters (for example        sodium dodecyl sulfate), lecithin, phospholipids, fatty alcohols        (such as, for example, Lanette®), sorbitan fatty acid esters        (such as, for example, Span®), polyoxyethylene sorbitan fatty        acid esters (such as, for example, Tween®), polyoxyethylene        fatty acid glycerides (such as, for example, Cremophor®),        polyoxethylene fatty acid esters, polyoxyethylene fatty alcohol        ethers, glycerol fatty acid esters, poloxamers (such as, for        example, Pluronic®),    -   buffers, acids and bases (for example phosphates, carbonates,        citric acid, acetic acid, hydrochloric acid, sodium hydroxide        solution, ammonium carbonate, trometamol, triethanolamine),    -   isotonicity agents (for example glucose, sodium chloride),    -   adsorbents (for example highly-disperse silicas),    -   viscosity-increasing agents, gel formers, thickeners and/or        binders (for example polyvinylpyrrolidone, methylcellulose,        hydroxypropylmethylcellulose, hydroxypropylcellulose,        carboxymethylcellulose-sodium, starch, carbomers, polyacrylic        acids (such as, for example, Carbopol®); alginates, gelatine),    -   disintegrants (for example modified starch,        carboxymethylcellulose-sodium, sodium starch glycolate (such as,        for example, Explotab®), cross-linked polyvinylpyrrolidone,        croscarmellose-sodium (such as, for example, AcDiSol®)),    -   flow regulators, lubricants, glidants and mould release agents        (for example magnesium stearate, stearic acid, talc,        highly-disperse silicas (such as, for example, Aerosil®)),    -   coating materials (for example sugar, shellac) and film formers        for films or diffusion membranes which dissolve rapidly or in a        modified manner (for example polyvinylpyrrolidones (such as, for        example, Kollidon®), polyvinyl alcohol,        hydroxypropylmethylcellulose, hydroxypropylcellulose,        ethylcellulose, hydroxypropylmethylcellulose phthalate,        cellulose acetate, cellulose acetate phthalate, polyacrylates,        polymethacrylates such as, for example, Eudragit®)),    -   capsule materials (for example gelatine,        hydroxypropylmethylcellulose),    -   synthetic polymers (for example polylactides, polyglycolides,        polyacrylates, polymethacrylates (such as, for example,        Eudragit®), polyvinylpyrrolidones (such as, for example,        Kollidon®), polyvinyl alcohols, polyvinyl acetates, polyethylene        oxides, polyethylene glycols and their copolymers and        blockcopolymers),    -   plasticizers (for example polyethylene glycols, propylene        glycol, glycerol, triacetine, triacetyl citrate, dibutyl        phthalate),    -   penetration enhancers,    -   stabilisers (for example antioxidants such as, for example,        ascorbic acid, ascorbyl palmitate, sodium ascorbate,        butylhydroxyanisole, butylhydroxytoluene, propyl gallate),    -   preservatives (for example parabens, sorbic acid, thiomersal,        benzalkonium chloride, chlorhexidine acetate, sodium benzoate),    -   colourants (for example inorganic pigments such as, for example,        iron oxides, titanium dioxide),    -   flavourings, sweeteners, flavour- and/or odour-masking agents.

The present invention furthermore relates to a pharmaceuticalcomposition which comprise at least one compound according to theinvention, conventionally together with one or more pharmaceuticallysuitable excipient(s), and to their use according to the presentinvention.

Based upon standard laboratory techniques known to evaluate compoundsuseful for the treatment of cardiovascular and renal disorders, bystandard toxicity tests and by standard pharmacological assays for thedetermination of treatment of the conditions identified above inmammals, and by comparison of these results with the results of knownactive ingredients or medicaments that are used to treat theseconditions, the effective dosage of the compounds of the presentinvention can readily be determined for treatment of each desiredindication. The amount of the active ingredient to be administered inthe treatment of one of these conditions can vary widely according tosuch considerations as the particular compound and dosage unit employed,the mode of administration, the period of treatment, the age and sex ofthe patient treated, and the nature and extent of the condition treated.

The total amount of the active ingredient to be administered willgenerally range from about 0.001 mg/kg to about 200 mg/kg body weightper day, and preferably from about 0.01 mg/kg to about 20 mg/kg bodyweight per day. Clinically useful dosing schedules will range from oneto three times a day dosing to once every four weeks dosing. Inaddition, it is possible for “drug holidays”, in which a patient is notdosed with a drug for a certain period of time, to be beneficial to theoverall balance between pharmacological effect and tolerability. It ispossible for a unit dosage to contain from about 0.5 mg to about 1500 mgof active ingredient, and can be administered one or more times per dayor less than once a day. The average daily dosage for administration byinjection, including intravenous, intramuscular, subcutaneous andparenteral injections, and use of infusion techniques will preferably befrom 0.01 to 200 mg/kg of total body weight. Illustratively, thecompound of the present invention may be administered parenterally at adose of about 0.001 mg/kg to about 10 mg/kg, preferably of about 0.01mg/kg to about 1 mg/kg of body weight. In oral administration, anexemplary dose range is about 0.01 to 100 mg/kg, preferably about 0.01to 20 mg/kg, and more preferably about 0.1 to 10 mg/kg of body weight.Ranges intermediate to the above-recited values are also intended to bepart of the invention.

Of course the specific initial and continuing dosage regimen for eachpatient will vary according to the nature and severity of the conditionas determined by the attending diagnostician, the activity of thespecific compound employed, the age and general condition of thepatient, time of administration, route of administration, rate ofexcretion of the drug, drug combinations, and the like. The desired modeof treatment and number of doses of a compound of the present inventionor a pharmaceutically acceptable salt or ester or composition thereofcan be ascertained by those skilled in the art using conventionaltreatment tests.

The following exemplary embodiments illustrate the invention. Theinvention is not restricted to the examples.

The percentages in the following tests and examples are, unless statedotherwise, by weight; parts are by weight. Solvent ratios, dilutionratios and concentrations reported for liquid/liquid solutions are eachbased on volume.

EXPERIMENTAL SECTION

NMR peak forms are stated as they appear in the spectra, possible higherorder effects have not been considered.

Chemical names were generated using the ACD/Name software from ACD/Labs.In some cases generally accepted names of commercially availablereagents were used in place of ACD/Name generated names.

The following table 1 lists the abbreviations used in this paragraph andin the Examples section as far as they are not explained within the textbody. Other abbreviations have their meanings customary per se to theskilled person.

TABLE 1 Abbreviations The following table lists the abbreviations usedherein. Abbreviation Meaning br broad (¹H-NMR signal) CI chemicalionisation d doublet (¹H-NMR signal) DCM dichloromethane dddouble-doublet (¹H-NMR signal) DMSO dimethylsulfoxide ESI electrospray(ES) ionisation h hour(s) HPLC high performance liquid chromatographyLC-MS liquid chromatography mass spectrometry m multiplet (¹H-NMRsignal) min minute(s) MS mass spectrometry NMR nuclear magneticresonance spectroscopy: chemical shifts (δ) are given in ppm. Thechemical shifts were corrected by setting the DMSO signal to 2.50 ppmunless otherwise stated. Rt retention time (as measured either with HPLCor UPLC) in minutes s singlet (¹H-NMR signal) SQDSingle-Quadrupole-Detector t triplet (¹H-NMR signal) td triple-doublet(¹H-NMR signal) THF tetrahydrofuran UPLC ultra performance liquidchromatography

The various aspects of the invention described in this application areillustrated by the following examples which are not meant to limit theinvention in any way.

The example testing experiments described herein serve to illustrate thepresent invention and the invention is not limited to the examplesgiven.

Experimental Section—General Part

All reagents, for which the synthesis is not described in theexperimental part, are either commercially available, or are knowncompounds or may be formed from known compounds by known methods by aperson skilled in the art.

The compounds and intermediates produced according to the methods of theinvention may require purification. Purification of organic compounds iswell known to the person skilled in the art and there may be severalways of purifying the same compound. In some cases, no purification maybe necessary. In some cases, the compounds may be purified bycrystallization. In some cases, impurities may be stirred out using asuitable solvent. In some cases, the compounds may be purified bychromatography, particularly flash column chromatography, using forexample prepacked silica gel cartridges, e.g. Biotage SNAP cartidgesKP-Sil® or KP-NH® in combination with a Biotage autopurifier system(SP4® or Isolera Four®) and eluents such as gradients of hexane/ethylacetate or DCM/methanol. In some cases, the compounds may be purified bypreparative HPLC using for example a Waters autopurifier equipped with adiode array detector and/or on-line electrospray ionization massspectrometer in combination with a suitable prepacked reverse phasecolumn and eluents such as gradients of water and acetonitrile which maycontain additives such as trifluoroacetic acid, formic acid or aqueousammonia.

In some cases, purification methods as described above can provide thosecompounds of the present invention which possess a sufficiently basic oracidic functionality in the form of a salt, such as, in the case of acompound of the present invention which is sufficiently basic, atrifluoroacetate or formate salt for example, or, in the case of acompound of the present invention which is sufficiently acidic, anammonium salt for example. A salt of this type can either be transformedinto its free base or free acid form, respectively, by various methodsknown to the person skilled in the art, or be used as salts insubsequent biological assays. It is to be understood that the specificform (e.g. salt, free base etc.) of a compound of the present inventionas isolated and as described herein is not necessarily the only form inwhich said compound can be applied to a biological assay in order toquantify the specific biological activity.

UPLC-MS Standard Procedures

Method 1 (LC/MS):

Instrument: Waters ACQUITY SQD UPLC System; Column: Waters Acquity UPLCHSS T3 1.8μ 50×1 mm; eluent A: 1 L water+0.25 ml 99% formic acid, eluentB: 1 L acetonitrile+0.25 ml 99% formic acid; gradient: 0.0 min 90% A→1.2min 5% A→2.0 min 5% A; oven: 50° C.; flow rate: 0.40 ml/min; UVdetection: 208-400 nm.

Method 2 (LC/MS):

Instrument MS: Thermo Scientific FT-MS; Instrument type UHPLC+: ThermoScientific UltiMate 3000; Column: Waters, HSST3, 2.1×75 mm, C18 1.8 μm;eluent A: 1 L water+0.01% formic acid; eluent B: 1 L acetonitrile+0.01%formic acid; gradient: 0.0 min 10% B→2.5 min 95% B→3.5 min 95% B; oven:50° C.; flow rate: 0.90 ml/min; UV detection: 210 nm/optimum integrationpath 210-300 nm.

Method 3 (LC/MS):

Instrument: Waters ACQUITY SQD UPLC System; Column: Waters Acquity UPLCHSS T3 1.8μ 50×1 mm; eluent A: 1 L water+0.25 ml 99% formic acid, EluentB: 1 L acetonitrile+0.25 ml 99% formic acid; gradient: 0.0 min 95% A→6.0min 5% A→7.5 min 5% A; oven: 50° C.; flow rate: 0.35 ml/min; UVdetection: 210-400 nm.

Method 4 (Preparative HPLC):

Column: Chromatorex or Reprosil C18 10 μm; 125×30 mm, Flow: 75 ml/min,Run time: 20 min, Detection at 210 nm, Eluent A: water+0.1% formic acid,Eluent B: acetonitrile+0.1% formic acid; Gradient: 3 min 10% B; 17.5min: 95% B; 19.5 min 100% B, 20 min 10% B.

EXPERIMENTAL SECTION—STARTING MATERIALS AND INTERMEDIATES Example 1A5-(4-Chlorophenyl)-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one

A solution of5-(4-chlorophenyl)-4-(3,3,3-trifluoro-2,2-dihydroxypropyl)-2,4-dihydro-3H-H-1,2,4-triazol-3-one(synthesis described as Example 3A in WO 2010/105770-A1) (10.0 g, 30.9mmol), N-[(1R,2R)-2-amino-1,2-diphenylethyl]-4-methylbenzenesulfonamide(56.6 mg, 154 μmol) and 1-methyl-4-(propan-2-yl)benzenedichlororuthenium (1:1) (47.3 mg, 77.2 μmol) in ethyl acetate wastreated with triethylamine (8.6 ml, 62 mmol) followed by addition offormic acid (5.8 ml, 150 mmol). The resulting mixture was heated underreflux for 3 h and then cooled down to room temperature. The reactionmixture was then diluted with 1N hydrochloric acid (70 ml). The organicphase was washed twice with 1N hydrochloric acid. The aqueous phase wasextracted twice with ethyl acetate. The combined organic phases wereevaporated. The residue was retaken in methanol (22.5 ml) and theresulting suspension was heated to 60° C. until the solid was completelydissolved. 1N hydrochloric acid (22.5 ml) was added and the resultingsuspension was heated at 78° C. for 10 min and then cooled down to roomtemperature. The solid was filtered off and dried under vacuum. Thesolid was retaken in iN hydrochloric acid (30 ml), heated at 35° C. Theresulting suspension was treated with methanol (30 ml), heated 4 h at35° C. and filtered off at 35° C. The filtrate solution was evaporatedaffording 4.9 g (ee=99.6%, 51% of th.) of5-(4-Chlorophenyl)-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one.

LC-MS (Method 2): R_(t)=1.40 min; MS (ESIpos): m/z=308 [M+H]⁺

¹H-NMR (400 MHz, DMSO): δ [ppm]=12.10 (s, 1H), 7.52-7.79 (m, 4H), 6.84(d, 1H), 3.54-4.52 (m, 3H).

Example 2A{3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetonitrile

In a 2 L reaction vessel, 100 g (273 mmol) of{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}aceticacid (synthesis described as Example 8A in WO 2010/105770-A1), 43.3 g(547 mmol) of pyridine and 33 mg (0.3 mmol) of 4-dimethyl-aminopyridinewere dissolved in 300 ml THF. The resulting solution was treated at 5°C. with 52.8 g (438 mmol) of 2,2-dimethylpropanoylchloride over 15minutes and the resulting mixture was stirred at room temperature for2.5 hours. After cooling to 0° C., 183 ml of 28% aqueous ammoniasolution was added over 1 h while the solution temperature was keptbetween 10° C. and 20° C. and at the resulting mixture then stirred at5° C. for an additional time period of 1 h. 500 ml methyltert-butylether and 300 ml 20% aqueous citric acid were then added whilekeeping the internal temperature between 10° C. and 20° C. The phaseswere the separated and the organic phase was washed with 300 ml of 20%aqueous citric acid followed by 300 ml saturated aqueous sodiumhydrogencarbonate solution and finally with 300 ml of 10% aqueous sodiumchloride solution. The organic phase was evaporated at 60° C. underreduced pressure until an oily residue was obtained. 300 ml THF was thenadded and the solution was evaporated again until an oily solution wasobtained. This operation was repeated a second time. The oil residue wasretaken in 360 ml THF and treated with 172 g (820 mmol) trifluoroaceticacid anhydride over 20 min at a temperature between 10° C. and 20° C.The resulting solution was then stirred at room temperature for 1 h. 720ml 4-methyl-2-pentanone and 650 ml 7.5% aqueous sodium hydroxidesolution were added at a temperature between 10° C. and 20° C. Finallythe pH-value was adjusted to pH=9.5 using 7.5% aqueous sodium hydroxidesolution. After phase separation, the organic phase was washed twicewith 450 ml 10% aqueous sodium chloride solution. The organic phase wasevaporated at a temperature of 80° C. under reduced pressure while 1200ml n-heptane was added. The formed suspension was cooled to 20° C. and asolid formed which was filtered off and washed with 200 ml n-heptane andthen dried under reduced pressure (50° C., 30 mbar) affording 88 g (93%of th.) of{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetonitrileas a solid.

1H-NMR (400 MHz, DMSO-d6): δ [ppm]=7.78 (d, 2H), 7.55 (d, 2H), 6.91 (d,1H), 5.17 (s, 2H), 4.34-4.23 (m, 1H), 3.98 (dd, 1H), 3.81 (dd, 1H).

Example 3A{3-(4-Chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetonitrile

A solution of 40 g (130 mmol)5-(4-Chlorophenyl)-4-[(2R)-3,3,3-trifluor-2-hydroxypropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one(Example 1A) in 400 ml methylisobutyl ketone was treated with 17.9 g(143 mmol) bromoacetonitrile and 53.9 g (390 mmol) potassium carbonateand stirred for 4 hours at 60° C. After cooling to 20° C., 200 ml waterwas added and the mixture was stirred for 10 min. After phaseseparation, the organic phase was washed with 200 ml water. The organicphase was evaporated at 80° C. under reduced pressure while 300 mln-heptane was added. The formed suspension was cooled to 20° C. and asolid formed which was filtered off and and washed with 50 ml n-heptaneand then dried under reduced pressure (50° C., 30 mbar) affording 25.2 g(56% of th.) of{3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetonitrile.

1H-NMR (400 MHz, DMSO-d6): δ [ppm]=7.78 (d, 2H), 7.55 (d, 2H), 6.91 (d,1H), 5.17 (s, 2H), 4.34-4.23 (m, 1H), 3.98 (dd, 1H), 3.81 (dd, 1H).

Example 4AMethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate

In a 4 L reaction vessel, 200 g (576.9 mmol) of{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetonitrile(Example 2A) in 1600 ml methanol was treated with 5.2 g (28 mmol) sodiummethanolate (30% in methanol) and the resulting mixture was stirred at50° C. for 2.5 hours. The solution was then evaporated at 50° C. underreduced pressure until an oily solution was obtained. 2000 ml methyltert-butylether was added and the solution was concentrated until avolume of 800 ml was achieved. 3000 ml n-heptane was then added and asuspension was formed. After cooling at 20° C., the solid was filteredand washed with 500 ml n-heptane and then dried under reduced pressure(50° C., 30 mbar) affording 175 g (80% of th.) of methyl2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate as a solid.

1H-NMR (400 MHz, DMSO-d6): δ [ppm]=8.01 (s, 1H), 7.78 (d, 2H), 7.62 (d,2H), 6.93 (br. s, 1H), 4.50 (s, 2H), 4.35-4.23 (m, 1H), 3.96 (dd, 1H),3.81 (dd, 1H), 3.67 (s, 3H).

Example 5AMethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate

A solution of 8.58 g (24.7 mmol) of{3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}acetonitrile(Example 3A) in methanol (43 ml) was treated with 229 μl (1.24 mmol) ofa sodium methoxide solution (30% in methanol). The resulting mixture wasstirred overnight at room temperature and then evaporated affording 9.31g (99% of th.) of the title compound.

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.01 (s, 1H), 7.81-7.58 (m, 4H),7.00-6.84 (m, 1H), 4.50 (s, 2H), 4.40-4.23 (m, 1H), 4.04-3.74 (m, 2H),3.66 (s, 3H).

Example 6A Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxylate

Under argon, a solution of 150 mg (0.40 mmol) ofmethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate(Example 4A) in 3 ml anhydrous THF was treated at 0° C. with 75 μl (0.44mmol) N,N-diisopropylethylamine and 40 μl (0.44 mmol) methylchlorooxoacetate and the mixture was then stirred at 0° C. for 30 min.93 mg (0.44 mmol) of [2-(trifluoromethyl)phenyl]hydrazine were thenadded, followed by 145 μl (0.83 mmol) of N,N-diisopropylethylamine. Theresulting mixture was stirred for 2 h at room temperature, followed by 1h at 120° C. in the microwave and then evaporated. The obtained residuewas purified by preparative HPLC (Method 4) affording 75 mg (32% of th.)of the title compound.

LC-MS (Method 2): R_(t)=2.01 min; MS(ESIpos): m/z=591.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.07-7.56 (m, 8H), 6.91 (d, 1H), 5.17(d, 2H), 4.37-4.21 (m, 1H), 4.09-3.80 (m, 2H), 3.74 (s, 3H).

Example 7A Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxylate

Under argon, a solution of 1 g (2.64 mmol) ofmethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate(Example 5A) in 3 ml anhydrous THF was treated at 0° C. with 506 μl(2.90 mmol) N,N-diisopropylethylamine and 267 μl (2.90 mmol) methylchlorooxoacetate and the mixture was then stirred at 0° C. for 30 min.617 mg (2.9 mmol) of [2-(trifluoromethyl)phenyl]hydrazine hydrochloridewere then added, followed by 506 μl (2.90 mmol) ofN,N-diisopropylethylamine. The resulting mixture was stirred for 2 h atroom temperature, followed by 1 h at 120° C. in the microwave and thenevaporated. The obtained residue was purified by flash chromatography(silica gel, eluent cyclohexane/ethyl acetate) followed by preparativeHPLC (Method 4) affording 485 mg (31% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.97 min; MS(ESIpos): m/z=591.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.03-7.57 (m, 8H), 6.91 (d, 1H), 5.17(d, 2H), 4.38-3.80 (m, 3H), 3.74 (s, 3H).

Example 8A Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethoxy)phenyl]-1H-1,2,4-triazole-5-carboxylate

Under argon, a solution of 150 mg (0.40 mmol) ofmethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate(Example 4A) in 3 ml anhydrous THF was treated at 0° C. with 75 μl (0.44mmol) N,N-diisopropylethylamine and 40 μl (0.44 mmol) methylchlorooxoacetate and the mixture was then stirred at 0° C. for 30 min.100 mg (0.44 mmol) of [2-(trifluoromethoxy)phenyl]hydrazinehydrochloride were then added, followed by 145 μl (0.83 mmol) ofN,N-diisopropylethylamine. The resulting mixture was stirred for 1.5 hat room temperature, followed by 1 h at 120° C. in the microwave andevaporated. The obtained residue was purified by preparative HPLC(Method 4) affording 129 mg (51% of th.) of the title compound.

LC-MS (Method 2): R_(t)=2.03 min; MS(ESIpos): m/z=607.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=7.86-7.52 (m, 8H), 6.91 (d, 1H), 5.18(d, 2H), 4.40-4.21 (m, 1H), 4.09-3.81 (m, 2H), 3.77 (s, 3H).

Example 9A Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethoxy)phenyl]-1H-1,2,4-triazole-5-carboxylate

Under argon, a solution of 1 g (2.64 mmol) ofmethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate(Example 5A) in 3 ml anhydrous THF was treated at 0° C. with 506 μl(2.90 mmol) N,N-diisopropylethylamine and 267 μl (2.90 mmol) methylchlorooxoacetate and the mixture was then stirred at 0° C. for 30 min.617 mg (2.9 mmol) of [2-(trifluoromethyl)phenyl]hydrazine hydrochloridewere then added, followed by 506 μl (2.90 mmol) ofN,N-diisopropylethylamine. The resulting mixture was stirred 1 h at roomtemperature, followed by 1 h at 120° C. in the microwave and thenevaporated. The obtained residue was purified by flash chromatography(silica gel, eluent cyclohexane/ethyl acetate) followed by preparativeHPLC (Method 4) affording 765 mg (45% of th.) of the title compound.

LC-MS (Method 2): R_(t)=2.02 min; MS(ESIpos): m/z=607.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm] 7.85-7.46 (m, 8H), 6.91 (d, 1H), 5.18(m, 2H), 4.36-3.79 (m, 3H), 3.77 (s, 3H).

Example 10A Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-(2-ethoxyphenyl)-1H-1,2,4-triazole-5-carboxylate

Under argon, a solution of 150 mg (0.40 mmol) ofmethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate(Example 4A) in 3 ml anhydrous THF was treated at 0° C. with 75 μl (0.44mmol) N,N-diisopropylethylamine and 40 μl (0.44 mmol) methylchlorooxoacetate and the mixture was then stirred at 0° C. for 30 min.82 mg (0.44 mmol) of [2-(ethyloxy)phenyl]hydrazine hydrochloride werethen added, followed by 145 μl (0.83 mmol) of N,N-diisopropylethylamine.The resulting mixture was stirred for 2 h at room temperature, followedby 1 h at 120° C. in the microwave and then evaporated. The obtainedresidue was purified by preparative HPLC (Method 4) affording 75 mg (33%of th.) of the title compound.

LC-MS (Method 1): R_(t)=1.09 min; MS(ESIpos): m/z=567.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=7.81-7.44 (m, 6H), 7.24-7.05 (m, 2H),6.91 (d, 1H), 5.15 (s, 2H), 4.39-4.21 (br m, 1H), 4.06-3.71 (m, 7H),1.13 (t, 3H).

Example 11A Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(methylcarbamoyl)phenyl]-1H-1,2,4-triazole-5-carboxylate

Under argon, a solution of 315 mg (0.83 mmol) ofmethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate(Example 4A) in 6.5 ml anhydrous THF was treated at 0° C. with 290 μl(1.66 mmol) N,N-diisopropylethylamine and 122 μl (1.66 mmol) methylchlorooxoacetate and the mixture was then stirred at 0° C. for 30 min.360 mg (0.91 mmol) of 2-hydrazinyl-N-methylbenzamide (synthesisdescribed in Journal of Heterocyclic Chemistry, 1979, 76(7), 1411) werethen added, followed by 290 μl (1.66 mmol) of N,N-diisopropylethylamine.The resulting mixture was stirred 1.5 h at room temperature, followed by1 h at 120° C. in the microwave and then evaporated. The obtainedresidue was purified by preparative HPLC (Method 4) affording 100 mg(69% of th.) of the title compound.

LC-MS (Method 3): R_(t)=2.71 min; MS(ESIpos): m/z=580.3 [M+H]⁺

Example 12A Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(methylsulfanyl)phenyl]-1H-1,2,4-triazole-5-carboxylate

Under argon, a solution of 300 mg (0.79 mmol) ofmethyl-2-{3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluor-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate(Example 4A) in 6 ml anhydrous THF was treated at 0° C. with 151 μl(0.44 mmol) N,N-diisopropylethylamine and 80 μl (0.87 mmol) methylchlorooxoacetate and the mixture was then stirred at 0° C. for 30 min.134 mg (0.87 mmol) of [2-(methylsulfanyl)phenyl]hydrazine was added,followed by 289 μl (1.66 mmol) of N,N-diisopropylethylamine. Theresulting mixture was stirred for 1.5 h at room temperature, followed by1 h at 120° C. in the microwave and then evaporated. The obtainedresidue was purified by preparative HPLC (Method 4) affording 325 mg(68% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.97 min; MS(ESIpos): m/z=569.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=7.88-7.25 (m, 8H), 6.92 (d, 1H), 5.16(s, 2H), 4.41-4.15 (m, 1H), 4.09-3.79 (m, 2H), 3.74 (s, 3H), 2.37 (s,3H).

Example 13A3-({3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(methylsulfanyl)phenyl]-1H-1,2,4-triazole-5-carboxamide

A mixture of 142 mg (0.25 mmol) of Example 12A in an ammonia solution(7N in methanol, 2 ml, 14 mmol) was stirred overnight at roomtemperature and the mixture was then evaporated. The obtained residuewas purified by preparative HPLC (Chromatorex C18, 10 μm, 125×30 mm,water/acetonitrile-gradient 0.1% formic acid) affording 107 mg (74% ofth.) of the title compound.

LC-MS (Method 2): R_(t)=1.72 min; MS(ESIpos): m/z=554.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.15 (s, 1H), 7.88-7.16 (m, 9H), 6.90(d, 1H), 5.13 (d, 2H), 4.40-4.19 (m, 1H), 4.08-3.75 (m, 2H), 2.35 (s,3H).

Example 14A Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-(2-ethoxyphenyl)-1H-1,2,4-triazole-5-carboxylate

Under argon, a solution of methyl2-{3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}ethanimidate(200 mg, 528 μmol, Example 5A) in anhydrous tetrahydrofurane (4.0 ml)was treated at 0° C. with N,N-diisopropylethylamine (100 μl, 580 μmol)and methyl chloro(oxo)acetate (53 μl, 580 μmol) and the mixture was thenstirred at 0° C. for 30 min. (2-Ethoxyphenyl)hydrazine hydrochloride(1:1) (110 mg, 581 μmol) was then added, followed byN,N-diisopropylethylamine (190 μl, 1.1 mmol). The resulting mixture wasstirred for 1.5 h at room temperature, followed by 1 h at 120° C. in themicrowave and then evaporated. The obtained residue was purified bypreparative HPLC (Method 4) affording 188 mg (61% of th.) of the titlecompound.

LC-MS (Method 2): R_(t)=1.98 min; MS (ESIpos): m/z=567.1 [M+H]⁺

¹H-NMR (400 MHz, DMSO-d6) δ[ppm]: 7.79-7.69 (m, 2H), 7.65-7.58 (m, 2H),7.50-7.46 (m, 2H), 7.21 (d, 1H), 7.10 (td, 1H), 6.91 (d, 1H), 5.15 (s,2H), 4.38-4.22 (m, 1H), 4.08-3.93 (m, 3H), 3.91-3.66 (m, 4H), 1.13 (t,3H).

EXPERIMENTAL SECTION—EXAMPLES Example 13-({3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxamide

A mixture of 55 mg (0.093 mmol) of methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxylate(Example 6A) in an ammonia solution (7N in methanol, 0.8 ml, 5.6 mmol)was stirred overnight at room temperature and the mixture was thenevaporated. The obtained residue was purified by preparative HPLC(Method 4) affording 55 mg (quant.) of the title compound.

LC-MS (Method 2): R_(t)=1.77 min; MS(ESIpos): m/z=576.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.23 (s, 1H), 8.00-7.55 (m, 9H), 6.90(d, 1H), 5.22-5.01 (m, 2H), 4.40-4.18 (m, 1H), 4.10-3.76 (m, 2H).

Example 23-({3-(4-Chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxamide

A mixture of 445 mg (0.75 mmol) of methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxylate(Example 7A) in an ammonia solution (7N in methanol, 10 ml, 70 mmol) wasstirred 1.5 h at room temperature and the mixture was then evaporatedaffording 398 mg (91% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.76 min; MS(ESIpos): m/z=576.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.23 (s, 1H), 7.99-7.55 (m, 9H), 6.90(d, 1H), 5.24-5.05 (m, 2H), 4.43-3.75 (m, 3H).

Example 33-({3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethoxy)phenyl]-1H-1,2,4-triazole-5-carboxamide

A mixture of 121 mg (0.20 mmol) of methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethoxy)phenyl]-1H-1,2,4-triazole-5-carboxylate(Example 8A) in an ammonia solution (7N in methanol, 1.7 ml, 11.9 mmol)was stirred overnight at room temperature and the mixture was thenevaporated. The obtained residue was purified by preparative HPLC(Method 4) affording 101 mg (86% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.82 min; MS(ESIpos): m/z=592.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.28 (s, 1H), 7.93 (s, 1H), 7.80-7.44(m, 8H), 6.90 (d, 1H), 5.25-4.97 (m, 2H), 4.44-4.19 (m, 1H), 4.07-3.76(m, 2H).

Example 43-({3-(4-Chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethoxy)phenyl]-1H-1,2,4-triazole-5-carboxamide

A mixture of 500 mg (0.82 mmol) of methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethoxy)phenyl]-1H-1,2,4-triazole-5-carboxylate(Example 9A) in an ammonia solution (7N in methanol, 10 ml, 70 mmol) wasstirred 2.5 h at room temperature and the mixture was then evaporatedaffording 479 mg (97% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.80 min; MS(ESIpos): m/z=592.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.28 (s, 1H), 7.93 (s, 1H), 7.80-7.46(m, 8H), 6.90 (d, 1H), 5.29-5.03 (m, 2H), 4.41-3.76 (m, 3H).

Example 53-({3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-(2-ethoxyphenyl)-1H-1,2,4-triazole-5-carboxamide

A mixture of 170 mg (0.30 mmol) of methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-(2-ethoxyphenyl)-1H-1,2,4-triazole-5-carboxylate(Example 10A) in an ammonia solution (7N in methanol, 3.2 ml, 22.4 mmol)was stirred overnight at room temperature and the mixture was thenevaporated. The obtained residue was purified by preparative HPLC(Method 4) affording 60 mg (36% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.76 min; MS(ESIpos): m/z=552.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.12 (s, 1H), 7.89-6.81 (m, 10H),5.11 (d, 2H), 4.39-4.20 (br m, 1H), 4.08-3.75 (m, 4H), 1.16 (t, 3H).

Example 63-({3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(methylcarbamoyl)phenyl]-1H-1,2,4-triazole-5-carboxamide

A mixture of 40 mg (70%, 0.05 mmol) of methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(methylcarbamoyl)-phenyl]-1H-1,2,4-triazole-5-carboxylate(Example 11A) in an ammonia solution (7N in methanol, 4 ml, 28 mmol) wasstirred overnight at room temperature and the mixture was thenevaporated. The obtained residue was purified by preparative HPLC(Method 4) affording 11 mg (43% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.47 min; MS(ESIpos): m/z=565.1 [M+H]⁺

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.33-7.34 (m, 11H), 6.92 (d, 1H),5.08 (s, 2H), 4.48-3.79 (m, 3H), 2.54 (s, 3H).

Example 73-({3-(4-Chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(methylsulfonyl)phenyl]-1H-1,2,4-triazole-5-carboxamide

At 0° C., a solution of 52 mg (0.20 mmol) of methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(methylsulfanyl)-phenyl]-1H-1,2,4-triazole-5-carboxylate(Example 13A) in 1 ml dichloromethane was treated with 69 mg (purity70%, 0.28 mmol) of 3-chloroperbenzoic acid and thereafter stirredovernight at room temperature. The reaction mixture was combined with afraction obtained from a 20 mg test run and the mixture was thenevaporated. The obtained residue was purified by preparative HPLC(Method 4) affording 60 mg (77% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.59 min; MS(ESIpos): m/z=586.1 [M+H]+

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.27-7.56 (m, 10H), 6.88 (d, 1H),5.16 (d, 2H), 4.37-4.12 (m, 1H), 4.07-3.73 (m, 2H), 3.07 (s, 3H).

Example 83-{[3-(4-chlorophenyl)-5-oxo-4-(3,3,3-trifluoro-2-oxopropyl)-4,5-dihydro-H-1,2,4-triazol-1-yl]methyl}-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxamide(ketone form) or3-{[3-(4-chlorophenyl)-5-oxo-4-(3,3,3-trifluoro-2,2-dihydroxypropyl)-4,5-dihydro-H-1,2,4-triazol-1-yl]methyl}-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxamide(hydrate form)

At 0° C., a solution of 150 mg (0.26 mmol) of3-({3-(4-chlorophenyl)-5-oxo-4-[(2S)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethyl)phenyl]-1H-1,2,4-triazole-5-carboxamide(Example 1) in 2.8 ml of dichloromethane was treated with 165 mg (0.39mmol) of Dess-Martin periodinane. The resulting mixture was stirred 1 hat 0° C. and then overnight at 4° C. 2 ml of saturated sodiumthiosulfate solution and 2 ml of a saturated sodium hydrogen carbonatesolution were then added and the resulting mixture was stirred for 10min at room temperature. The reaction mixture was extracted with ethylacetate (three times repeated), the combined organic layers were thenevaporated and the obtained residue purified by flash chromatography(silica gel, eluent cyclohexane/ethyl acetate) affording 122 mg (82% ofth.) of the title compound.

LC-MS (Method 2): R_(t)=1.78 min; MS(ESIpos): m/z=574.1 [M+H]⁺ (ketoneform)

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.24 (s, 1H), 7.96-7.53 (m, 9H), 7.44(s, 2H), 5.27-4.96 (m, 2H), 4.05 (s, 2H), (hydrate form).

Example 93-{[3-(4-chlorophenyl)-5-oxo-4-(3,3,3-trifluoro-2-oxopropyl)-4,5-dihydro-H-1,2,4-triazol-1-yl]methyl}-1-[2-(trifluoromethoxy)phenyl]-1H-1,2,4-triazole-5-carboxamide(ketone form) or3-{[3-(4-chlorophenyl)-5-oxo-4-(3,3,3-trifluoro-2,2-dihydroxypropyl)-4,5-dihydro-1H-1,2,4-triazol-1-yl]methyl}-1-[2-(trifluoromethoxy)phenyl]-1H-1,2,4-triazole-5-carboxamide(hydrate form)

At 0° C., a solution of 150 mg (0.26 mmol) of3-({3-(4-Chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-[2-(trifluoromethoxy)-phenyl]-1H-1,2,4-triazole-5-carboxamide(Example 4) in 2.8 ml of dichloromethane was treated with 165 mg (0.39mmol) of Dess-Martin periodinane. The resulting mixture was stirred 1 hat 0° C. and then a further 45 min at room temperature. 2 ml ofsaturated sodium thiosulfate solution and 2 ml of a saturated sodiumhydrogen carbonate solution were then added and the resulting mixturewas stirred for 10 min at room temperature. The reaction mixture wasextracted with dichloromethane (four times repeated). The combinedorganic layers were then evaporated and the obtained residue purified byflash chromatography (silica gel, eluent cyclohexane/ethyl acetate)affording 66 mg (44% of th.) of the title compound.

LC-MS (Method 2): R_(t)=1.80 min; MS(ESIpos): m/z=590 [M+H]⁺ (ketoneform)

¹H NMR (DMSO-d₆, 400 MHz): δ [ppm]=8.29 (s, 1H), 7.93 (s, 1H), 7.76-7.48(m, 8H), 7.44 (s, 2H), 5.16 (s, 2H), 4.05 (s, 2H) (hydrate form).

Example 103-({3-(4-Chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-H-1,2,4-triazol-1-yl}methyl)-1-(2-ethoxyphenyl)-1H-1,2,4-triazole-5-carboxamide

Methyl3-({3-(4-chlorophenyl)-5-oxo-4-[(2R)-3,3,3-trifluoro-2-hydroxypropyl]-4,5-dihydro-1H-1,2,4-triazol-1-yl}methyl)-1-(2-ethoxyphenyl)-1H-1,2,4-triazole-5-carboxylate(143 mg, 253 μmol, Example 14A) in ammonia solution in methanol (2.7 ml,7n) was stirred overnight at room temperature and evaporated. Theresidue was evaporated and purified by preparative HPLC (Method 4)affording 118 mg (84% of th.) of the title compound.

LC-MS (Method 1): R_(t)=0.93 min; MS (ESIpos): m/z=552.1 [M+H]⁺

¹H-NMR (400 MHz, DMSO-d6) δ[ppm]: 8.12 (s, 1H), 7.87-7.71 (m, 3H),7.66-7.57 (m, 2H), 7.47-7.33 (m, 2H), 7.15 (d, 1H), 7.08-6.99 (m, 1H),6.91 (d, 1H), 5.22-4.99 (m, 2H), 4.41-4.19 (m, 1H), 4.11-3.74 (m, 4H),1.16 (t, 3H).

EXPERIMENTAL SECTION—BIOLOGICAL ASSAYS Abbreviations and Acronyms

Acc. No. accession number AVP arginine vasopressin B_(max) maximalligand binding capacity BSA bovine serum albumin cAMP cyclic adenosinemonophosphate Cat. No. catalogue number cDNA complementarydeoxyribonucleic acid CHO chinese hamster ovary CRE cAMP responseelement Ct cycle threshold DMEM/F12 Dulbecco's modified Eagle'smedium/Ham's F12 medium (1:1) DNA deoxyribonucleic acid DMSOdimethylsulfoxide DTT dithiothreitol EC₅₀ half-maximal effectiveconcentration EDTA ethylenediamine-tetraacetic acid FAMcarboxyfluorescein succinimidyl ester f.c. final concentration FCS fetalcalf serum HEPES 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid IC₅₀half-maximal inhibitory concentration K_(d) dissociation constant K_(i)dissociation constant of an inhibitor mRNA messenger ribonucleic acidPBS phosphate buffered saline PEG polyethylene glycol p.o. per os,peroral RNA ribonucleic acid RTPCR real-time polymerase chain reactionSPA scintillation proximity assay TAMRA carboxytetramethylrhodamineTRIS; Tris 2-amino-2-hydroxymethylpropane-1,3-diol

Examples were tested in selected biological assays one or more times.When tested more than once, data are reported as either average valuesor as median values, wherein

-   -   the average value, also referred to as the arithmetic mean        value, represents the sum of the values obtained divided by the        number of times tested, and    -   the median value represents the middle number of the group of        values when ranked in ascending or descending order. If the        number of values in the data set is odd, the median is the        middle value. If the number of values in the data set is even,        the median is the arithmetic mean of the two middle values.

Examples were synthesized one or more times. When synthesized more thanonce, data from biological assays represent average values or medianvalues calculated utilizing data sets obtained from testing of one ormore synthetic batch.

Demonstration of the activity of the compounds of the present inventionmay be accomplished through in vitro, ex vivo, and in vivo assays thatare well known in the art. For example, to demonstrate the activity ofthe compounds of the present invention, the following assays may beused.

B-1. Cellular In Vitro Assay for Determining Vasopressin ReceptorActivity

The identification of agonists and antagonists of the V1a and V2vasopressin receptors from humans, rats and dogs as well as thequantification of the activity of the compounds of the invention iscarried out using recombinant cell lines. These cell lines originallyderive from a hamster's ovary epithelial cell (Chinese Hamster Ovary,CHO K1, ATCC: American Type Culture Collection, Manassas, Va. 20108,USA). The test cell lines constitutively express the human, rat or dogV1a or V2 receptors. In case of the G_(αq)-coupled V1a receptors, cellsare also stably transfected with a modified form of thecalcium-sensitive photoproteins aequorin (human and rat V1a) or obe-lin(dog V1a), which, after reconstitution with the cofactor coelenterazine,emit light when there are increases in free calcium concentrations[Rizzuto R, Simpson A W, Brini M, Pozzan T, Nature 358, 325-327 (1992);Illarionov B A, Bondar V S, Illarionova V A, Vysotski E S, Gene 153 (2),273-274 (1995)]. The resulting vasopressin receptor cells react tostimulation of the recombinantly expressed V1a receptors byintracellular release of calcium ions, which can be quantified by theresulting photoprotein luminescence. The G_(s)-coupled V2 receptors arestably transfected into cell lines expressing the gene for fireflyluciferase under control of a CRE-responsible promoter. Activation of V2receptors induces the activation of the CRE-responsive promoter via cAMPincrease, thereby inducing the expression of firefly luciferase. Thelight emitted by photoproteins of V1a cell lines as well as the lightemitted by firefly luciferase of V2 cell lines corresponds to theactivation or inhibition of the respective vasopressin receptor. Thebioluminescence of the cell lines is detected using a suitableluminometer [Milligan G, Marshall F, Rees S, Trends in PharmacologicalSciences 17, 235-237 (1996)].

Test Procedure:

Vasopressin V1a Receptor Cell Lines:

On the day before the assay, the cells are plated out in culture medium(DMEM/F12, 2% FCS, 2 mM glutamine, 10 mM HEPES, 5 μg/ml coelenterazine)in 384-well microtiter plates and kept in a cell incubator (96%humidity, 5% v/v CO₂, 37° C.). On the day of the assay, test compoundsin various concentrations are placed for 10 minutes in the wells of themicrotiter plate before the agonist [Arg⁸]-vasopressin at EC₅₀concentration is added. The resulting light signal is measuredimmediately in a luminometer.

Vasopressin V2 Receptor Cell Lines:

On the day before the assay, the cells are plated out in culture medium(DMEM/F12, 2% FCS, 2 mM glutamine, 10 mM HEPES) in 384-well microtiterplates and kept in a cell incubator (96% humidity, 5% v/v CO₂, 37° C.).On the day of the assay, test compounds in various concentrations andthe agonist [Arg⁸]-vasopressin at EC₅₀ concentration are added togetherto the wells, and plates are incubated for 3 hours in a cell incubator.Upon addition of the cell lysis reagent Triton™ and the substrateluciferin, luminescence of firefly luciferase is measured in aluminometer.

Table 1A below lists individual IC₅₀ values for the compounds of theinvention (including racemic mixtures as well as separated enantiomers)that were obtained from cell lines transfected with the human V1a or V2receptor:

TABLE 1A Example IC₅₀ hV1a IC₅₀ hV2 ratio IC₅₀ No. [μM] [μM] hV2/hV1a 10.00058 0.02390 41.4 2 0.00235 0.17500 74.5 3 0.00125 0.07550 60.4 40.00965 0.66500 68.9 5 0.00160 0.03850 24.1 6 0.01290 0.97500 75.6 70.01750 0.71250 40.7 8 0.00185 0.03367 18.2 9 0.00895 0.10650 11.9 100.00820 0.89000 108.5

The IC₅₀ data listed in Table 1A demonstrate that the compounds of thepresent invention are acting as selective and potent vasopressin V1areceptor antagonists.

For comparative purposes, selected phenyl-triazole derivatives that wereregarded to be representative of closest prior art (cf Int. Pat. Appl.WO 2011/104322-A1 and example compounds described therein) were alsotested in the cellular V1a and V2 assays described above. IC₅₀ valuesfor these compounds obtained from cell lines transfected with the humanV1a or V2 receptor are listed in Table 1B below:

TABLE 1B Example No. IC₅₀ hV1a IC₅₀ hV2 ratio IC₅₀ WO 2011/104322 [μM][μM] hV2/hV1a 63 0.0068 0.0042 0.622 64 0.0329 0.0345 1.049 66 1.82650.0950 0.052 67 2.4650 1.1400 0.462 99 0.1393 0.2184 1.568 143 0.45900.9090 1.98

For comparative purposes, further selected phenyl-triazole derivativesthat were regarded to be representative of closest prior art (cf Int.Pat. Appl. WO 2016/071212-A1 and example compounds described therein)were also tested in the cellular V1a and V2 assays described above. IC₅₀values for these compounds obtained from cell lines transfected with thehuman V1a or V2 receptor are listed in Table 1C below:

TABLE 1C Example No. IC₅₀ hV1a IC₅₀ hV2 ratio IC₅₀ WO 2016/071212 [μM][μM] hV2/hV1a 8 0.0012 0.0107 8.78 73 0.0011 0.0070 6.48 74 0.00220.0247 11.44

B-2. Radioactive Binding Assay

IC₅₀ and K_(i) values can be determined in radioactive binding assaysusing membrane fractions of recombinant human embryonic kidney cell line293 (HEK293) or CHO-K1 cell lines expressing the respective humanvasopressin V1a and V2 receptors.

Human recombinant vasopressin V1a receptors expressed in HEK293 cellsare used in 50 mM Tris-HCl buffer, pH 7.4, 5 mM MgCl₂, 0.1% BSA usingstandard techniques. Aliquots of prepared membranes are incubated withtest compounds in various concentrations in duplicates and 0.03 nM[¹²⁵I]Phenylacetyl-D-Tyr(Me)-Phe-Gln-Asn-Arg-Pro-Arg-Tyr-NH₂ for 120minutes at 25° C. Non-specific binding is estimated in the presence of 1μM [Arg⁸]Vasopressin. Receptors are filtered and washed, the filters arethen counted to determine[¹²⁵I]Phenylacetyl-D-Tyr(Me)-Phe-Gln-Asn-Arg-Pro-Arg-Tyr-NH₂specifically bound.

CHO-K1 cells stably transfected with a plasmid encoding humanvasopressin V2 receptor are used to prepare membranes in 50 mM Tris-HClbuffer, pH 7.4, 10 mM MgCl₂, 0.1% BSA using standard techniques.Aliquots of prepared membrane are incubated with test compounds invarious concentrations in duplicates and 4 nM [³H](Arg⁸)-Vasopressin for120 minutes at 25° C. Non-specific binding is estimated in the presenceof 1 mM (Arg⁸)-vasopressin. Membranes are filtered and washed 3 timesand the filters are counted to determine [³H](Arg₈)-Vasopressinspecifically bound.

IC₅₀ values are determined by a non-linear, least squares regressionanalysis using MathIQ™ (ID Business Solutions Ltd., UK). The inhibitionconstant K_(i) is calculated using the equation of Cheng and Prusoff(Cheng, Y., Prusoff, W. H., Biochem. Pharmacol. 22:3099-3108, 1973).

B-3. Cellular In Vitro Assay for Detecting the Action of Vasopressin V1aReceptor Antagonists on the Regulation of Pro-Fibrotic Genes

The cell line H9C2 (American Type Culture Collection ATCC No. CRL-1446),described as a cardiomyocyte type isolated from rat cardiac tissue,endogenously expresses the vasopressin V1a receptor AVPR1A in high copynumber, whereas AVPR2 expression cannot be detected. Likewise, the cellline NRK49F (ATCC No. CRL1570) isolated from rat kidney tissue, showssimilar expression pattern of high AVPR1A mRNA expression anddiminishing AVPR2 expression. For cell assays detecting the inhibitionof AVPR1A receptor-dependent regulation of gene expression by receptorantagonists, the procedure is as follows:

H9C2 cells or NRK49F cells are seeded in 6-well microtiter plates forcell culture at a cell density of 50 000 cells/well in 2.0 ml ofOpti-MEM medium (Invitrogen Corp., Carlsbad, Calif., USA, Cat. No.11058-021) and held in a cell incubator (96% humidity, 8% v/v CO₂, 37°C.). After 24 hours, sets of three wells (triplicate) are charged withvehicle solution (negative control) and vasopressin solution([Arg8]-vasopressin acetate, Sigma, Cat. No. V9879), or test compound(dissolved in vehicle: water with 20% v/v ethanol) and vasopressinsolution. In the cell culture, the final vasopressin concentration is 1nM. The test compound solution is added to the cell culture in smallvolumes, so that a final concentration of 0.03% of ethanol in the cellassay is not exceeded. After an incubation time of 5 hours, the culturesupernatant is drawn off under suction, the adherent cells are lysed in350 μl of RLT buffer (Qiagen, Cat. No. 79216), and the RNA is isolatedfrom the lysate using the RNeasy kit (Qiagen, Cat. No. 74104). This isfollowed by DNAse digestion (Invitrogen, Cat. No. 18068-015), cDNAsynthesis (Promaga, ImProm-II Reverse Transcription System, Cat. No.A3800) and Reverse Transcription Polymerase Chain Reaction (RTPCR) (pPCRMasterMix RT-QP2X-03-075, Eurogentec, Seraing, Belgium). All procedurestake place in accordance with the working protocols of the testreagents' manufacturers. The primer sets for the RTPCR are selected onthe basis of the mRNA gene sequences (NCBI GenBank Entrez NucleotideData Base) using the Primer3Plus program with 6-FAM TAMRA-labelledprobes. The RTPCR for determining the relative mRNA expression in thecells of the various assay batches is carried out using the AppliedBiosystems ABI Prism 7700 Sequence Detector in 384-well microtiter plateformat in accordance with the instrument operating instructions. Therelative gene expression is represented by the delta-delta Ct value[Applied Biosystems, User Bulletin No. 2 ABI Prism 7700 SDS, Dec. 11,1997 (updated October 2001)] with reference to the level of expressionof the ribosomal protein L-32 gene (GenBank Acc. No. NM_013226) and thethreshold Ct value of Ct=35.

B-4. Inhibition of Vasopressin Induced Aggregation of Human Platelets

Human platelets endogenously express the V1a receptor. It was found thatrelatively high vasopressin concentrations (ca. 50-100 nM) stimulateplatelet aggregation ex vivo. Therefore, platelets enriched from humanblood may serve as a V1a expressing tissue for pharmacological studieswith corresponding high concentrations of vasopressin antagonists.

Human blood is collected in a 10 mM trisodium citrate solution by venouspuncture from nonsmoking healthy volunteers (n=4-8) who were drug freefor at least 1 week. Platelet-rich plasma (PRP) is obtained bycentrifuging the blood sample at 140 g for 20 min at 4° C. The resultingpellet is further centrifuged (15.000 rpm, 2 min) to produceplatelet-poor plasma (PPP). Platelet aggregation is measuredturbidimetrically using an aggregometer (APACT 4). The reaction isfollowed by monitoring changes in light transmission on 178 μL PRPaliquots, under continuous stirring at 37° C., against PPP control.Various concentrations of vasopressin antagonists (in 2 μL) are added toPRP 5 min before the addition of 20 μL Arg-vasopressin (finalconcentration 100 nM. The inhibitory effects of the compounds aredetermined by measuring the height of the aggregation wave from thebottom of the shape change compared with the control response. IC50values are calculated a dose-response inhibition curve by an iterativenonlinear regression program

B-5. Effects on the Contraction of Isolated Rat Vessel Rings

Isolated Aorta

Test compounds can be investigated on isolated aortic rings from maleWistar rats endogenously expressing the V1a receptor. Male Wistar ratsare euthanized using carbon dioxide. The aorta is removed and placed inice-cold Krebs-Henseleit buffer of following composition (in mmol/l):NaCl 112, KCl 5.9, CaCl₂ 2.0, MgCl₂ 1.2, NaH₂PO₄ 1.2, NaHCO₃ 25, glucose11.5. The aorta is cut into 3 mm rings and transferred to 20 ml organbaths containing Krebs-Henseleit solution equilibrated with 95% O₂, 5%CO₂ at 37° C. For recording of isometric tension the rings are mountedbetween two hooks. The resting tension is adjusted to 3 g. After anequilibration period, each experiment is started by exposing thepreparation to K+(50 mM) Krebs-Henseleit solution. The aortic rings arethan pre-contracted using 1 nmol/l Arg-vasopressin. After a stablecontraction is established, a cumulative dose response curve of the testcompound is constructed. The stabilized contraction induced byArg-vasopressin is defined as 100% tension. The relaxation is expressedas percentage tension.

Isolated A. Renalis

Male Wistar rats (200-250 g) are euthanized using carbon dioxide. The A.renalis is removed and placed in ice-cold Krebs-Henseleit buffer offollowing composition (in mmol/1): NaCl 112, KCl 5.9, CaCl₂ 2.0, MgCl₂1.2, NaH₂PO₄ 1.2, NaHCO₃ 25, glucose 11.5. For measurement of isometrictension, ring segments, 2 mm in length, are mounted in a small vesselchamber myograph (Danish Myo Technology A/S, Denmark) using two tungstenwires fixed to mounting jaws. One mounting jaw is attached to amicrometer, allowing control of vessel circumference. The other mountingjaw is attached to a force transducer for measurement of tensiondevelopment. The whole preparation is kept in a chamber withphysiological salt solution at 37° C., bubbled with oxygen. After a 30min equilibration period, the vessels are stretched to their optimallumen diameter for active tension development which is determined basedon the internal circumference-wall tension ratio. The internalcircumference is set to 90% of what the vessels would have if they areexposed to a passive tension equivalent to that produced by a transmuralpressure of 100 mmHg.

Afterwards, the vessels are washed three times with Krebs-Henseleitbuffer and left to equilibrate for 30 min. The contractility is thentested by a twofold exposure to a high K⁺ solution (50 mmol/l KCl).After washing with Krebs-Henseleit buffer the vessels are thenpre-contracted using 1 nmol/l Arg-vasopressin. After a stablecontraction is established, a cumulative dose response curve of the testcompound is constructed. The stabilized contraction induced byArg-vasopressin is defined as 100% tension. The relaxation is expressedas percentage tension.

B-6. In Vivo Assay for Detecting Cardiovascular Effects: Blood PressureMeasurement in Anaesthetized Rats (Vasopressin ‘Challenge’ Model)

Male Sprague-Dawley rats (250-350 g body weight) are used underketamine/xylazine/pentobarbital injection anaesthesia. Polyethylenetubes (PE-50, Intramedic®), prefilled with heparin-containing (500IU/ml) isotonic sodium chloride solution, are introduced into thejugular vein and the femoral vein and then tied in. Arg-vasopressin(SIGMA) is injected via one venous access, with the aid of a syringe;the test substance is administered via the second venous access. Fordetermination of the systolic blood pressure, a pressure catheter(Millar SPR-320 2F) is tied into the carotid artery. The arterialcatheter is connected to a pressure transducer which feeds its signalsto a recording computer equipped with suitable recording software. In atypical experiment, the experimental animal is administered 3-4successive bolus injections at intervals of 10-15 min with a definedamount of Arg-vasopressin (30 ng/kg) in isotonic sodium chloridesolution. When the blood pressure has reached initial levels again, thetest substance is administered as a bolus, with subsequent continuousinfusion, in a suitable solvent. After this, at defined intervals (10-15min), the same amount of Arg-vasopressin as at the start is administeredagain. On the basis of the blood pressure values, a determination ismade of the extent to which the test substance counteracts thehypertensive effect of Arg-vasopressin. Control animals only receivesolvent instead of the test substance.

Following intravenous administration, the compounds of the invention, incomparison to the solvent controls, bring about an inhibition of theblood pressure increase caused by Arg-vasopressin.

B-7. In Vivo Assay for Detecting Protective Renal Effects: AcuteIschemia/Reperfusion Injury Model in Rodents

Laboratory bred male C57Bl/6J mice 6-8 weeks old are obtained fromTaconic Biosciences, male 6-8 weeks old Sprague Dawley® rat are obtainedfrom Charles River. Both rats and mice are maintained under standardlaboratory conditions, 12 hour light-dark cycles with access to normalchow and drinking water at libitum. For the ischemia reperfusion injurymodel a total of 10-12 rats or mice is used in each control andexperimental group.

Animals are anesthetized with continuous inhaled isoflurane. A rightnephrectomy is performed through a right flank incision 7 days beforethe ischemic procedures in the contralateral kidneys. For renal ischemiaa left flank incision is made. Renal vessels are exposed by dissectionof the left renal pedicle. Non-traumatic vascular clamps are used tostop blood flow (artery and vein) during 45 min (rats) or 25 min (mice)of ischemia. Reperfusion is established by removing the clamps. Theabdominal wall (muscular layer and skin) is closed with 5.0polypropylene sutures. Temgesic® (Buprenorphin, 0.025 mg/kg s.c.) isapplied as an analgesic.

Urine of each animal is collected in metabolic cages over night beforesacrifice at 24 h post ischemia. Upon sacrifice, blood samples areobtained under terminal anesthesia. After centrifugation of the bloodsamples, serum is isolated. Both serum creatinine and serum urea aremeasured via clinical biochemistry analyzer (Pentra 400). For theassessment of serum and urinary kidney injury biomarkers (Neutrophilgelatinase-associated lipocalin [NGAL], kidney injury molecule-1 [KIM-1]and Osteopontin) ELISA's are performed according to the manufacturersprotocol. Both urinary creatinine and albumin are measured to determinethe albumin/creatinine ratio.

Total RNA is isolated from kidneys. Left kidneys are snap-frozen inliquid nitrogen at sacrifice. Kidney tissue is then homogenized and RNAis obtained. Total RNA is transcribed to cDNA. Using TaqMan real-timePCR renal NGAL, Osteopontin, KIM-1, Nephrin and Podocin mRNA expressionis analyzed in whole kidney tissue.

Differences between groups are analyzed by one-way ANOVA with Dunnett'scorrections for multiple comparisons. Statistical significance isdefined as p<0.05. All statistical analyses are done using GraphPadPrism 6.

B-8. In Vivo Assay for Detecting Cardiovascular Effects: HemodynamicInvestigations in Anaesthetized Dogs

Male beagle dogs (Beagle, Marshall BioResources, USA) with a weight ofbetween 10 and 15 kg are anesthetized with pentobarbital (30 mg/kg iv,Narcoren®, Merial, Germany) for the surgical interventions and thehemodynamic and functional investigation termini. Pancuroniumbromide(Pancuronium Inresa, Inresa, Germany, 2-4 mg/animal i.v.) servesadditionally as a muscle relaxant. The dogs are intubated and ventilatedwith an oxygen/ambient air mixture (30/70%), about 2,5-4 L/min.Ventilation takes place using a ventilator from GE Healthcare (Avance,Germany) and is monitored using a carbon dioxide analyzer (-DatexOhmeda). The anesthesia is maintained by continual infusion ofpentobarbital (50 μg/kg/min); fentanyl is used as an analgesic (10μg/kg/h).

In preparatory interventions, the dogs are fitted with a cardiacpacemaker. At start of experiment, a cardiac pacemaker from Biotronik(Logos®, Germany) is implanted into a subcutaneous skin pocket and iscontacted with the heart via a pacemaker electrode (Siello S60®,Biotronik, Germany) which is advanced through the external jugular vein,with illumination, into the right ventricle.

Thereafter accesses are removed and the dog wakes spontaneously from theanesthesia. After a further 7 days, the above-described pacemaker isactivated and the heart is stimulated at a frequency of 220 beats perminute.

The actual drug testing experiments take place 28 days after thebeginning of pacemaker stimulation, using the following instrumentation:

-   -   Introduction of a bladder catheter for bladder relief and for        measuring the flow of urine    -   Attachment of electrocardiography (ECG) leads to the extremities        for ECG measurement    -   Introduction of a sheath introducer filled with sodium chloride        solution into the femoral artery. This tube is connected to a        pressure sensor (Braun Melsungen, Melsungen, Germany) for        measuring the systemic blood pressure    -   Introduction of a Millar Tip catheter (type 350 PC, Millar        Instruments, Houston, USA) through a port secured in the carotid        artery, for measuring cardiac hemodynamics.    -   Introduction of a Swan-Ganz catheter (CCOmbo 7.5F, Edwards,        Irvine, USA) via the jugular vein into the pulmonary artery, for        measuring the cardiac output, oxygen saturation, pulmonary        arterial pressures and central venous pressure    -   Siting of a venous catheter in the cephalic vein, for infusing        pentobarbital, for liquid replacement and for blood sampling        (determination of the plasma levels of substance or other        clinical blood values)    -   Siting of a venous catheter in the saphenous vein, for infusing        fentanyl and for administration of substance    -   Infusion of vasopressin (Sigma) in increasing dosage, up to a        dose of 4 mU/kg/min. The pharmacological substances are then        tested with this dosage.

The primary signals are amplified if necessary (ACQ7700, Data SciencesInternational, USA or Edwards-Vigilance-Monitor, Edwards, Irvine, USA)and subsequently fed into the Ponemah system (Data SciencesInternational, USA) for evaluation. The signals are recordedcontinuously throughout the experimental period, and are furtherprocessed digitally by said software, and averaged over 30 seconds.

B-9. Determination of Pharmacokinetic Parameters Following Intravenousand Oral Administration

The pharmacokinetic parameters of the compounds according to theinvention are determined in male C57b16-mice, male Wistar rats, femaleBeagle dogs and female Cynomolgus monkeys. Intravenous administration inthe case of mice and rats is carried out by means of a species-specificplasma/DMSO formulation, and in the case of dogs and monkeys by means ofa water/PEG400/ethanol formulation. In all species, oral administrationof the dissolved substance is performed via gavage, based on awater/PEG400/ethanol formulation. The taking of blood from rats issimplified by inserting a silicone catheter into the right Venajugularis externa prior to substance administration. The operation iscarried out at least one day prior to the experiment with isoflurananaesthesia and administration of an analgesic (atropin/Rimadyl (3/1)0.1 ml s.c.). The blood is taken (generally at least 10 time points)within a time window including terminal time points of at least 24 to amaximum of 72 hours after substance administration. When the blood istaken, it is passed into heparinised tubes. Then the blood plasma isobtained by centrifugation and is optionally stored at −20° C. untilfurther processing.

An internal standard (which may also be a chemically unrelatedsubstance) is added to the samples of the compounds according to theinvention, calibration samples and qualifiers, and there follows proteinprecipitation by means of excess acetonitrile. Addition of a buffersolution matched to the LC conditions, and subsequent vortexing, isfollowed by centrifugation at 1000 g. The supernatant is analysed byLC-MS/MS using C18 or biphenyl reversed-phase columns and variablemobile phase mixtures. The substances are quantified via the peakheights or areas from extracted ion chromatograms of specific selectedion monitoring experiments.

The plasma concentration/time plots determined are used to calculate thepharmacokinetic parameters such as AUC (area under the curve), C_(max)(maximal concentration), t_(1/2) (terminal half-life), F(bioavailability), MRT (mean residence time) and CL (clearance), using avalidated pharmacokinetic calculation program.

Since the substance quantification is carried out in plasma, it isnecessary to determine the blood/plasma distribution of the substance inorder to be able to adjust the pharmacokinetic parameterscorrespondingly. For this purpose, a defined amount of substance isincubated in heparinized whole blood of the species in question in arocking roller mixture for 20 min. After centrifugation at 1000 g, theplasma concentration is measured (by means of LC-MS/MS; see above) anddetermined by calculating the ratio of the whole blood concentrationversus plasma concentration (C_(blood)/C_(plasma) Value).

B-10. Metabolic Study

To determine the metabolic profile of the compounds according to theinvention, they are incubated with recombinant human cytochrome P450(CYP) enzymes, liver microsomes or primary fresh hepatocytes fromvarious animal species (e.g. rats, dogs, monkeys), and also of humanorigin, in order to obtain and to compare information aboutsubstantially the complete hepatic phase I and phase II metabolism, andabout the enzymes involved in the metabolism.

The compounds according to the invention were incubated with aconcentration of about 0.1-10 μM. To this end, stock solutions of thecompounds according to the invention having a concentration of 0.01-1 mMin acetonitrile were prepared, and then pipetted with 1:100 dilutioninto the incubation mixture. The liver microsomes and recombinantenzymes were incubated at 37° C. in 50 mM potassium phosphate buffer pH7.4 with and without Nicotinamide adenine dinucleotide phosphate(NADPH)-generating system consisting of 1 mM NADP⁺, 10 mM glucose6-phosphate and 1 unit of glucose 6-phosphate dehydrogenase. Primaryhepatocytes were incubated in suspension in Williams E medium, likewiseat 37° C. After an incubation time of 0-4 h, the incubation mixtureswere stopped with acetonitrile (final concentration about 30%), and theprotein was centrifuged off at about 15 000×g. The samples thus stoppedwere either analysed directly or stored at −20° C. until analysis.

The analysis is carried out by means of high-performance liquidchromatography with ultraviolet and mass spectrometry detection(HPLC-UV-MS/MS). To this end, the supernatants of the incubation samplesare chromatographed with suitable C18 reversed-phase columns andvariable mobile phase mixtures of acetonitrile and 10 mM aqueousammonium formate solution or 0.05% formic acid. The UV chromatograms inconjunction with mass spectrometry data serve for identification,structural elucidation and quantitative estimation of the metabolites,and for quantitative metabolic assessment of the compound according tothe invention in the incubation mixtures.

B-11. Caco-2 Permeability Test

The permeability of a test substance can be determined with the aid ofthe Caco-2 cell line, an established in vitro model for permeabilitypredictions at the gastrointestinal barrier (Artursson, P. and Karlsson,J. (1991). Correlation between oral drug absorption in humans andapparent drug permeability coefficients in human intestinal epithelial(Caco-2) cells. Biochem. Biophys. 175 (3), 880-885). The CaCo-2 cells(ACC No. 169, DSMZ, Deutsche Sammlung von Mikroorganismen undZellkulturen, Braunschweig, Germany) are seeded in 24-well plates withinset and cultivated for 14 to 16 days. For the permeability studies,the test substance is dissolved in DMSO and diluted with transportbuffer (Hanks Buffered Salt Solution, Gibco/Invitrogen, with 19.9 mMglucose and 9.8 mM HEPES) to the final test concentration. To determinethe permeability from the apical to the basolateral side (P_(app)A-B) ofthe test substance, the solution comprising the test substance is placedon the apical side of the Caco-2 cell monolayer, and the transportbuffer on the basolateral side. To determine the permeability from thebasolateral to the apical side (P_(app)B-A) of the test substance, thesolution comprising the test substance is placed on the basolateral sideof the Caco-2 cell monolayer, and the transport buffer on the apicalside. At the start of the experiment, samples are taken from therespective donor compartment to calculate the mass balance afterwards.After a two-hour incubation at 37° C., samples are taken from the twocompartments. The samples are analysed by LC-MS/MS, and the apparentpermeability coefficients (P_(app)) are calculated. For each cellmonolayer, the permeability of Lucifer Yellow is determined to ensurecell layer integrity. In each test run, the permeability of atenolol(marker for low permeability) and sulfasalazine (marker for activeexcretion) is also determined as quality control.

C) Working Examples of Pharmaceutical Compositions

The substances according to the invention can be converted topharmaceutical preparations as follows:

Tablet:

Composition:

100 mg of the compound of Example 1, 50 mg of lactose (monohydrate), 50mg of maize starch, 10 mg of polyvinylpyrrolidone (PVP 25) (from BASF,Germany) and 2 mg of magnesium stearate.

Tablet weight 212 mg. Diameter 8 mm, radius of curvature 12 mm.

Production:

The mixture of the compound of Example 1, lactose and starch isgranulated with a 5% strength solution (m/m) of the PVP in water. Afterdrying, the granules are mixed with the magnesium stearate for 5 min.This mixture is compressed in a conventional tabletting press (see abovefor format of the tablet).

Oral Suspension:

Composition:

1000 mg of the compound of Example 1, 1000 mg of ethanol (96%), 400 mgof Rhodigel (xanthan gum) (from FMC, USA) and 99 g of water.

10 ml of oral suspension correspond to a single dose of 100 mg of thecompound of the invention.

Production:

The Rhodigel is suspended in ethanol, and the compound of Example 1 isadded to the suspension.

The water is added while stirring. The mixture is stirred for about 6 huntil swelling of the Rhodigel is complete.

Sterile i.v. Solution:

The compound according to the invention is dissolved at a concentrationbelow saturation solubility in a physiologically acceptable solvent (forexample isotonic sodium chloride solution, glucose solution 5% and/orPEG 400 solution 30%). The solution is sterilized by filtration andfilled into sterile and pyrogen-free injection containers.

Although the invention has been disclosed with reference to specificembodiments, it is apparent that other embodiments and variations of theinvention may be devised by others skilled in the art without departingfrom the true spirit and scope of the invention. The claims are intendedto be construed to include all such embodiments and equivalentvariations.

1. A compound of general formula (I)

in which R¹ represents a group of the formula

in which #¹ represents the point of attachment to the nitrogen atom, Arrepresents a group of the formula

in which #² represents the point of attachment to the nitrogen atom, R²represents a group selected from trifluoromethyl, trifluoromethoxy,ethoxy, —C(═O)N(H)CH₃ and —S(═O)₂CH₃, or a pharmaceutically acceptablesalt, hydrate and/or solvate thereof.
 2. The compound of general formula(I) according to claim 1, wherein R¹ represents a group of the formula

in which #¹ represents the point of attachment to the nitrogen atom, R²represents a group selected from trifluoromethyl, trifluoromethoxy,ethoxy, —C(═O)N(H)CH₃ and —S(═O)₂CH₃, or a pharmaceutically acceptablesalt, hydrate and/or solvate thereof.
 3. The compound of general formula(I) according to claim 1, wherein R¹ represents a group of the formula

in which #¹ represents the point of attachment to the nitrogen atom, R²represents a group selected from trifluoromethyl and trifluoromethoxy,or a pharmaceutically acceptable salt, hydrate and/or solvate thereof.4. A method of preparing a compound of general formula (I) according toclaim 1 said method comprising the step [A] of allowing an intermediatecompound of formula (II):

in which R¹ is as defined for the compound of general formula (I)according to claim 1, R³ represents a (C₁-C₄)-alkyl group, in particulara methyl group, to react in a first step in the presence of a base witha compound of general formula (III):

in which R⁴ represents a (C₁-C₄)-alkyl group, in particular a methylgroup, to give an intermediate compound, which is then allowed to reactin the presence of a base in a second step with a hydrazine compound ofgeneral formula (IV) or a respective salt thereof

in which Ar is as defined for the compound of general formula (I)according to claim 1, thereby giving a compound of general formula (V):

in which R¹ and Ar are as defined for the compound of general formula(I) according to claim 1, and R⁴ represents a (C₁-C₄)-alkyl group, inparticular a methyl group, followed by a subsequent step [B] of allowingthe compound of formula (V) obtained in step [A] to react with ammoniathereby giving a compound of general formula (I):

in which R¹ and Ar are as defined for the compound of general formula(I) according to claim 1, optionally followed by step [C] conversion ofthe alcohols of general formula (I-A):

in which Ar is as defined for the compound of general formula (I)according to claim 1, to the ketones of general formula (I-B):

in which Ar is as defined for the compound of general formula (I)according to claim 1, using known oxidation methods, each [A], [B] and[C] optionally followed, where appropriate, by (i) separating thecompounds of formula (I) thus obtained into their respectiveenantiomers, and/or (ii) converting the compounds of formula (I) intotheir respective hydrates, solvates, salts and/or hydrates or solvatesof the salts by treatment with the corresponding solvents and/or acidsor bases.
 5. The compound as defined in claim 1 for use in the treatmentand/or prevention of a diseases.
 6. The compound as defined in claim 1for use in a method for the treatment and/or prevention of acute kidneydisease, chronic kidney disease, diabetic nephropathy, acute heartfailure, chronic heart failure, preeclampsia, peripheral arterialdisease (PAD), coronary microvascular dysfunction (CMD), Raynaud'ssyndrome, and dysmenorrhea.
 7. A method of manufacturing apharmaceutical composition for the treatment and/or prevention of adisease selected from the group consisting of acute kidney disease,chronic kidney disease, diabetic nephropathy, acute heart failure,chronic heart failure, preeclampsia, peripheral arterial disease (PAD),coronary microvascular dysfunction (CMD), Raynaud's syndrome, anddysmenorrhea; the method comprising manufacturing the pharmaceuticalcomposition with a compound of claim
 1. 8. Pharmaceutical compositioncomprising a compound as defined in claim 1 and one or morepharmaceutically acceptable excipients.
 9. Pharmaceutical compositioncomprising one or more first active ingredients, in particular compoundsof general formula (I) according to claim 1, and one or more furtheractive ingredients.
 10. The pharmaceutical composition as defined inclaim 8 for the treatment and/or prevention of a disease selected fromthe group consisting of acute kidney disease, chronic kidney disease,diabetic nephropathy, acute heart failure, chronic heart failure,preeclampsia, peripheral arterial disease (PAD), coronary microvasculardysfunction (CMD), Raynaud's syndrome, and dysmenorrhea.
 11. Method forthe treatment and/or prevention of a disease selected from the groupconsisting of acute kidney disease, chronic kidney disease, diabeticnephropathy, acute heart failure, chronic heart failure, preeclampsia,peripheral arterial disease (PAD), coronary microvascular dysfunction(CMD), Raynaud's syndrome, and dysmenorrhea, in a human or other mammal,comprising administering to a human or other mammal in need thereof atherapeutically effective amount of one or more compounds as defined inclaim
 1. 12. Method for the treatment and/or prevention of a diseaseselected from the group consisting of acute kidney disease, chronickidney disease, diabetic nephropathy, acute heart failure, chronic heartfailure, preeclampsia, peripheral arterial disease (PAD), coronarymicrovascular dysfunction (CMD), Raynaud's syndrome, and dysmenorrhea,in a human or other mammal, comprising administering to a human or othermammal in need thereof a therapeutically effective amount of apharmaceutical composition as defined in claim
 8. 13. The pharmaceuticalcomposition of claim 9 wherein the one or more further activeingredients is selected from the group consisting of diuretic,angiotensin AII antagonist, ACE inhibitor, beta-receptor blocker,mineralocorticoid receptor antagonist, antidiabetic, organic nitrate, NOdonor, activator, and stimulator of the soluble guanylate cyclase,antiinflammatory agents, immunosuppressive agent, phosphate binder, andcompound which modulate vitamin D metabolism.